Inbred maize line PH4CV

ABSTRACT

An inbred maize line, designated PH4CV, the plants and seeds of inbred maize line PH4CV, methods for producing a maize plant, either inbred or hybrid, produced by crossing the inbred maize line PH4CV with another maize plant, and hybrid maize seeds and plants produced by crossing the inbred line PH4CV with another maize line or plant and to methods for producing a maize plant containing in its genetic material one or more transgenes and to the transgenic maize plants produced by that method. This invention also relates to inbred maize lines derived from inbred maize line PH4CV, to methods for producing other inbred maize lines derived from inbred maize line PH4CV and to the inbred maize lines derived by the use of those methods.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims benefit of the priority date of U.S. PatentApplication Ser. No. 60/352,317 filed Jan. 28, 2002, which is herebyincorporated herein by reference.

FIELD OF THE INVENTION

This invention is in the field of maize breeding, specifically relatingto an inbred maize line designated PH4CV.

BACKGROUND OF THE INVENTION

The goal of plant breeding is to combine in a single variety or hybrid,various desirable traits. For field crops, these traits may includeresistance to diseases and insects, tolerance to heat and drought,reducing the time to crop maturity, greater yield, and better agronomicquality. With mechanical harvesting of many crops, uniformity of plantcharacteristics such as germination and stand establishment, growthrate, maturity, and plant and ear height, is important.

Field crops are bred through techniques that take advantage of theplant's method of pollination. A plant is self-pollinated if pollen fromone flower is transferred to the same or another flower of the sameplant. A plant is sib-pollinated when individuals within the same familyor line are used for pollination. A plant is cross-pollinated if thepollen comes from a flower on a different plant from a different familyor line. The terms “cross-pollination” and “out-cross” as used herein donot include self-pollination or sib-pollination.

Plants that have been self-pollinated and selected for type for manygenerations become homozygous at almost all gene loci and produce auniform population of true breeding progeny. A cross between twodifferent homozygous lines produces a uniform population of hybridplants that may be heterozygous for many gene loci. A cross of twoplants, each heterozygous at a number of gene loci will produce apopulation of hybrid plants that differ genetically and will not beuniform.

Maize (zea mays L.), often referred to as corn in the United States, canbe bred by both self-pollination and cross-pollination techniques. Maizehas separate male and female flowers on the same plant, located on thetassel and the ear, respectively. Natural pollination occurs in maizewhen wind blows pollen from the tassels to the silks that protrude fromthe tops of the ears.

A reliable method of controlling male fertility in plants offers theopportunity for improved plant breeding. This is especially true fordevelopment of maize hybrids, which relies upon some sort of malesterility system. There are several ways in which a maize plant can bemanipulated so that it is male sterile. These include use of manual ormechanical emasculation (or detasseling), use of cytoplasmic genetic ornuclear genetic male sterility, use of gametocides and is the like.

Hybrid maize seed is typically produced by a male sterility systemincorporating manual or mechanical detasseling. Alternate strips of twomaize inbreds are planted in a field, and the pollen-bearing tassels areremoved from one of the inbreds (female). Provided that there issufficient isolation from sources of foreign maize pollen, the ears ofthe detasseled inbred will be fertilized only from the other inbred(male), and the resulting seed is therefore hybrid and will form hybridplants.

The laborious detasseling process can be avoided by using cytoplasmicmale-sterile (CMS) inbreds. Plants of a CMS inbred are male sterile as aresult of factors resulting from the cytoplasmic, as opposed to thenuclear, genome. Thus, this characteristic is inherited exclusivelythrough the female parent in maize plants, since only the femaleprovides cytoplasm to the fertilized seed. CMS plants are fertilizedwith pollen from another inbred that is not male-sterile. Pollen fromthe second inbred may or may not contribute genes that make the hybridplants male-fertile. The same hybrid seed, a portion produced fromdetasseled fertile maize and a portion produced using the CMS system,can be blended to insure that adequate pollen loads are available forfertilization when the hybrid plants are grown.

There are several methods of conferring genetic male sterilityavailable, such as multiple mutant genes at separate locations withinthe genome that confer male sterility, as disclosed in U.S. Pat. Nos.4,654,465 and 4,727,219 to Brar et al. and chromosomal translocations asdescribed by Patterson in U.S. Pat. Nos. 3,861,709 and 3,710,511. Theseand all patents, patent applications and publications referred to hereinare incorporated by reference. In addition to these methods, Albertsenet al., of Pioneer Hi-Bred, U.S. Pat. No. 5,432,068, have developed asystem of nuclear male sterility which includes: identifying a genewhich is critical to male fertility; silencing this native gene which iscritical to male fertility; removing the native promoter from theessential male fertility gene and replacing it with an induciblepromoter; inserting this genetically engineered gene back into theplant; and thus creating a plant that is male sterile because theinducible promoter is not “on” resulting in the male fertility gene notbeing transcribed. Fertility is restored by inducing, or turning “on”,the promoter, which in turn allows the gene that confers male fertilityto be transcribed.

These, and the other methods of conferring genetic male sterility in theart, each possess their own benefits and drawbacks. Some other methodsuse a variety of approaches such as delivering into the plant a geneencoding a cytotoxic substance associated with a male tissue specificpromoter or an antisense system in which a gene critical to fertility isidentified and an antisense to that gene is inserted in the plant (see:Fabinjanski, et al. EPO 89/3010153.8 publication no. 329,308 and PCTapplication PCT/CA90/00037 published as WO 90/08828).

Another system for controlling male sterility makes use of gametocides.Gametocides are not a genetic system, but rather a topical applicationof chemicals. These chemicals affect cells that are critical to malefertility. The application of these chemicals affects fertility in theplants only for the growing season in which the gametocide is applied(see Carlson, Glenn R., U.S. Pat. No. 4,936,904). Application of thegametocide, timing of the application and genotype specificity oftenlimit the usefulness of the approach and it is not appropriate in allsituations.

Development of Maize Inbred Lines

The use of male sterile inbreds is but one factor in the production ofmaize hybrids. Plant breeding techniques known in the art and used in amaize plant breeding program include, but are not limited to, recurrentselection, backcrossing, pedigree breeding, restriction fragment lengthpolymorphism enhanced selection, genetic marker enhanced selection,making double haploids, and transformation. Often a combination of thesetechniques is used. The development of maize hybrids in a maize plantbreeding program requires, in general, the development of homozygousinbred lines, the crossing of these lines, and the evaluation of thecrosses.

Maize plant breeding programs combine the genetic backgrounds from twoor more inbred lines or various other germplasm sources into breedingpopulations from which new inbred lines are developed by selfing andselection of desired phenotypes. The new inbreds are crossed with otherinbred lines and the hybrids from these crosses are evaluated todetermine which of those have commercial potential. Plant breeding andhybrid development, as practiced in a maize plant breeding programdeveloping significant genetic advancement, are expensive andtime-consuming processes.

Pedigree breeding starts with the crossing of two genotypes, such as twoelite inbred lines, each of which may have one or more desirablecharacteristics that is lacking in the other or which complements theother. If the two original parents do not provide all the desiredcharacteristics, other sources can be included in the breedingpopulation. In the pedigree method, superior plants are selfed andselected in successive filial generations. In the succeeding filialgenerations the heterozygous condition gives way to homogeneous lines asa result of self-pollination and selection. Typically in the pedigreemethod of breeding, five or more successive filial generations ofselfing and selection is practiced: F₁→F₂; F₂→F₃; F₃→F₄; F₄→F₅, etc.After a sufficient amount of inbreeding, successive filial generationswill serve to increase seed of the developed inbred. Preferably, aninbred line comprises homozygous alleles at about 95% or more of itsloci.

Backcrossing can be used to improve an inbred line and a hybrid that ismade using those inbreds. Backcrossing can be used to transfer aspecific desirable trait from one line, the donor parent, to an inbredcalled the recurrent parent which has overall good agronomiccharacteristics yet lacks that desirable trait. This transfer of thedesirable trait into an inbred with overall good agronomiccharacteristics can be accomplished by first crossing a recurrent parentand a donor parent (non-recurrent parent). The progeny of this cross isthen mated back to the recurrent parent followed by selection in theresultant progeny for the desired trait to be transferred from thenon-recurrent parent as well as selection for the characteristics of therecurrent parent. Typically after four or more backcross generationswith selection for the desired trait and the characteristics of therecurrent parent, the progeny will contain essentially all genes of therecurrent parent except for the genes controlling the desired trait.However, the number of backcross generations can be less if molecularmarkers are used during selection or elite germplasm is used as thedonor parent. The last backcross generation is then selfed to give purebreeding progeny for the gene(s) being transferred. Backcrossing canalso be used in conjunction with pedigree breeding to develop new inbredlines. For example, an F1 can be created that is backcrossed to one ofits parent lines to create a BC1, BC2, BC3, etc. Progeny are selfed andselected so that the newly developed inbred has many of the attributesof the recurrent parent and some of the desired attributes of thenon-recurrent parent. This approach leverages the value and strengths ofthe recurrent parent for use in new hybrids and breeding which has verysignificant value for a breeder.

Recurrent selection is a method used in a plant breeding program toimprove a population of plants. The method entails individual plantscross-pollinating with each other to form progeny, which are then grown.The superior progeny are then selected by any number of methods, whichinclude individual plant, half-sib progeny, full-sib progeny, selfedprogeny and topcrossing. The selected progeny are cross-pollinated witheach other to form progeny for another population. This population isplanted and again superior plants are selected to cross-pollinate witheach other. Recurrent selection is a cyclical process and therefore canbe repeated as many times as desired. The objective of recurrentselection is to improve the traits of a population. The improvedpopulation can then be used as a source of breeding material to obtaininbred lines to be used in hybrids or used as parents for a syntheticcultivar. A synthetic cultivar is the resultant progeny formed by theintercrossing of several selected inbreds. Mass selection is a usefultechnique when used in conjunction with molecular marker enhancedselection.

Mutation breeding is one of the many methods of introducing new traitsinto inbred lines. Mutations that occur spontaneously or areartificially induced can be useful sources of variability for a plantbreeder. The goal of artificial mutagenesis is to increase the rate ofmutation for a desired characteristic. Mutation rates can be increasedby many different means including temperature, long-term seed storage,tissue culture conditions, radiation; such as X-rays, Gamma rays (e.g.cobalt 60 or cesium 137), neutrons, (produce of nuclear fission byuranium 235 on an atomic reactor), Beta radiation (emitted fromradioisotopes such as phosphorus 32 or carbon 14), or ultravioletradiation (preferably from 2500 to 2900 nm), or chemical mutagens (suchas base analogues (5-bromo-uracil), related compounds (8-ethoxycaffeine), antibiotics (streptonigrin), alkylating agents (sulfurmustards, nitrogen mustards, epoxides, ethylenamines, sulfates,sulfonates, sulfones, lactones), azide, hydroxylamine, nitrous acid, oracridines. Once a desired trait is observed through mutagenesis thetrait may then be incorporated into existing germplasm by traditionalbreeding techniques. Details of mutation breeding can be found in“Principals of Cultivar Development” Fehr, 1993 Macmillan PublishingCompany the disclosure of which is incorporated herein by reference.

Molecular markers, which includes markers identified through the use oftechniques such as Isozyme Electrophoresis, Restriction Fragment LengthPolymorphisms (RFLPs), Randomly Amplified Polymorphic DNAs (RAPDs),Arbitrarily Primed Polymerase Chain Reaction (AP-PCR), DNA AmplificationFingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs),Amplified Fragment Length Polymorphisms (AFLPs), Simple Sequence Repeats(SSRs) and Single Nucleotide Polymorphisms (SNPs), may be used in plantbreeding methods. One use of molecular markers is Quantitative TraitLoci (QTL) mapping. QTL mapping is the use of markers, which are knownto be closely linked to alleles that have measurable effects on aquantitative trait. Selection in the breeding process is based upon theaccumulation of markers linked to the positive effecting alleles and/orthe elimination of the markers linked to the negative effecting allelesfrom the plant's genome.

Molecular markers can also be used during the breeding process for theselection of qualitative traits. For example, markers closely linked toalleles or markers containing sequences within the actual alleles ofinterest can be used to select plants that contain the alleles ofinterest during a backcrossing breeding program. The markers can also beused to select for the genome of the recurrent parent and against thegenome of the donor parent. Using this procedure can minimize the amountof genome from the donor parent that remains in the selected plants. Itcan also be used to reduce the number of crosses back to the recurrentparent needed in a backcrossing program. The use of molecular markers inthe selection process is often called genetic marker enhanced selection.

The production of double haploids can also be used for the developmentof inbreds in the breeding program. Double haploids are produced by thedoubling of a set of chromosomes (1N) from a heterozygous plant toproduce a completely homozygous individual. For example, see Wan et al.,“Efficient Production of Doubled Haploid Plants Through ColchicineTreatment of Anther-Derived Maize Callus”, Theoretical and AppliedGenetics, 77:889-892, 1989. This can be advantageous because the processomits the generations of selfing needed to obtain a homozygous plantfrom a heterozygous source.

Development of Maize Hybrids

A single cross maize hybrid results from the cross of two inbred lines,each of which has a genotype that complements the genotype of the other.The hybrid progeny of the first generation is designated F₁. In thedevelopment of commercial hybrids in a maize plant breeding program,only the F₁ hybrid plants are sought. F₁ hybrids are more vigorous thantheir inbred parents. This hybrid vigor, or heterosis, can be manifestedin many polygenic traits, including increased vegetative growth andincreased yield.

The development of a maize hybrid in a maize plant breeding programinvolves three steps: (1) the selection of plants from various germplasmpools for initial breeding crosses; (2) the selfing of the selectedplants from the breeding crosses for several generations to produce aseries of inbred lines, which, although different from each other, breedtrue and are highly uniform; and (3) crossing the selected inbred lineswith different inbred lines to produce the hybrids. During theinbreeding process in maize, the vigor of the lines decreases. Vigor isrestored when two different inbred lines are crossed to produce thehybrid. An important consequence of the homozygosity and homogeneity ofthe inbred lines is that the hybrid between a defined pair of inbredswill always be the same. Once the inbreds that give a superior hybridhave been identified, the hybrid seed can be reproduced indefinitely aslong as the homogeneity of the inbred parents is maintained.

A single cross hybrid is produced when two inbred lines are crossed toproduce the F₁ progeny. A double cross hybrid is produced from fourinbred lines crossed in pairs (A×B and C×D) and then the two F₁ hybridsare crossed again (A×B)×(C×D). A three-way cross hybrid is produced fromthree inbred lines where two of the inbred lines are crossed (A×B) andthen the resulting F₁ hybrid is crossed with the third inbred (A×B)×C.Much of the hybrid vigor and uniformity exhibited by F₁ hybrids is lostin the next generation (F₂). Consequently, seed produced from hybrids isnot used for planting stock.

Hybrid seed production requires elimination or inactivation of pollenproduced by the female parent. Incomplete removal or inactivation of thepollen provides the potential for self-pollination. This inadvertentlyself-pollinated seed may be unintentionally harvested and packaged withhybrid seed. Also, because the male parent is grown next to the femaleparent in the field there is the very low probability that the maleselfed seed could be unintentionally harvested and packaged with thehybrid seed. Once the seed from the hybrid bag is planted, it ispossible to identify and select these self-pollinated plants. Theseself-pollinated plants will be genetically equivalent to one of theinbred lines used to produce the hybrid. Though the possibility ofinbreds being included hybrid seed bags exists, the occurrence is verylow because much care is taken to avoid such inclusions. It is worthnoting that hybrid seed is sold to growers for the production of grainor forage and not for breeding or seed production.

These self-pollinated plants can be identified and selected by oneskilled in the art due to their decreased vigor when compared to thehybrid. Inbreds are identified by their less vigorous appearance forvegetative and/or reproductive characteristics, including shorter plantheight, small ear size, ear and kernel shape, cob color, or othercharacteristics.

Identification of these self-pollinated lines can also be accomplishedthrough molecular marker analyses. See, “The Identification of FemaleSelfs in Hybrid Maize: A Comparison Using Electrophoresis andMorphology”, Smith, J. S. C. and Wych, R. D., Seed Science andTechnology 14, pp. 1-8 (1995), the disclosure of which is expresslyincorporated herein by reference. Through these technologies, thehomozygosity of the self-pollinated line can be verified by analyzingallelic composition at various loci along the genome. Those methodsallow for rapid identification of the invention disclosed herein. Seealso, “Identification of Atypical Plants in Hybrid Maize Seed byPostcontrol and Electrophoresis” Sarca, V. et al., Probleme de GeneticaTeoritica si Aplicata Vol. 20 (1) p. 29-42.

As is readily apparent to one skilled in the art, the foregoing are onlysome of the various ways by which the inbred can be obtained by thoselooking to use the germplasm. Other means are available, and the aboveexamples are illustrative only.

Maize is an important and valuable field crop. Thus, a continuing goalof plant breeders is to develop high-yielding maize hybrids that areagronomically sound based on stable inbred lines. The reasons for thisgoal are obvious: to maximize the amount of grain produced with theinputs used and minimize susceptibility of the crop to pests andenvironmental stresses. To accomplish this goal, the maize breeder mustselect and develop superior inbred parental lines for producing hybrids.This requires identification and selection of genetically uniqueindividuals that occur in a segregating population. The segregatingpopulation is the result of a combination of crossover events plus theindependent assortment of specific combinations of alleles at many geneloci that results in specific genotypes. The probability of selectingany one individual with a specific genotype from a breeding cross isinfinitesimal due to the large number of segregating genes and theunlimited recombinations of these genes, some of which may be closelylinked. However, the genetic variation among individual progeny of abreeding cross allows for the identification of rare and valuable newgenotypes. These new genotypes are neither predictable nor incrementalin value, but rather the result of manifested genetic variation combinedwith selection methods, environments and the actions of the breeder.Once identified, it is possible to utilize routine and predictablebreeding methods to develop progeny that retain the rare and valuablenew genotypes developed by the initial breeder.

Even if the entire genotypes of the parents of the breeding cross werecharacterized and a desired genotype known, only a few if anyindividuals having the desired genotype may be found in a largesegregating F₂ population. It would be very unlikely that a breeder ofordinary skill in the art would able to recreate the same line twicefrom the very same original parents, as the breeder is unable to directhow the genomes combine or how they will interact with the environmentalconditions. This unpredictability results in the expenditure of largeamounts of research resources in the development of a superior new maizeinbred line. Once such a line is developed its value to society issubstantial since it is important to advance the germplasm base as awhole in order to maintain or improve traits such as yield, diseaseresistance, pest resistance and plant performance in extreme conditions.

A breeder uses various methods to help determine which plants should beselected from the segregating populations and ultimately which inbredlines will be used to develop hybrids for commercialization. In additionto the knowledge of the germplasm and other skills the breeder uses, apart of the selection process is dependent on experimental designcoupled with the use of statistical analysis. Experimental design andstatistical analysis are used to help determine which plants, whichfamily of plants, and finally which inbred lines and hybrid combinationsare significantly better or different for one or more traits ofinterest. Experimental design methods are used to assess error so thatdifferences between two inbred lines or two hybrid lines can be moreaccurately determined. Statistical analysis includes the calculation ofmean values, determination of the statistical significance of thesources of variation, and the calculation of the appropriate variancecomponents. Either a five or a one percent significance level iscustomarily used to determine whether a difference that occurs for agiven trait is real or due to the environment or experimental error.

One of ordinary skill in the art of plant breeding would know how toevaluate the traits of two plant varieties to determine if there is nosignificant difference between the two traits expressed by thosevarieties. For example, see Fehr, Walt, Principles of CultivarDevelopment, p. 261-286 (1987) which is incorporated herein byreference. Mean trait values may be used to determine whether traitdifferences are significant, and preferably the traits are measured onplants grown under the same environmental conditions.

Combining ability of a line, as well as the performance of the line perse, is a factor in the selection of improved maize inbreds. Combiningability refers to a line's contribution as a parent when crossed withother lines to form hybrids. The hybrids formed for the purpose ofselecting superior lines are designated testcrosses. One way ofmeasuring combining ability is by using breeding values. Breeding valuesare based in part on the overall mean of a number of testcrosses. Thismean is then adjusted to remove environmental effects and it is adjustedfor known genetic relationships among the lines.

SUMMARY OF THE INVENTION

According to the invention, there is provided a novel inbred maize line,designated PH4CV. This invention thus relates to the seeds of inbredmaize line PH4CV, to the plants of inbred maize line PH4CV, to plantparts of inbred maize line PH4CV, to methods for producing a maize plantproduced by crossing the inbred maize line PH4CV with another maizeplant, including a plant that is part of a synthetic or naturalpopulation, and to methods for producing a maize plant containing in itsgenetic material one or more transgenes and to the transgenic maizeplants and plant parts produced by that method. This invention alsorelates to inbred maize lines and plant parts derived from inbred maizeline PH4CV, to methods for producing other inbred maize lines derivedfrom inbred maize line PH4CV and to the inbred maize lines and theirparts derived by the use of those methods. This invention furtherrelates to hybrid maize seeds, plants, and plant parts produced bycrossing the inbred line PH4CV with another maize line.

Definitions

Certain definitions used in the specification are provided below. Alsoin the examples that follow, a number of terms are used herein. In orderto provide a clear and consistent understanding of the specification andclaims, including the scope to be given such terms, the followingdefinitions are provided. NOTE: ABS is in absolute terms and % MN ispercent of the mean for the experiments in which the inbred or hybridwas grown. PCT designates that the trait is calculated as a percentage.% NOT designates the percentage of plants that did not exhibit a trait.For example, STKLDG % NOT is the percentage of plants in a plot thatwere not stalk lodged. These designators will follow the descriptors todenote how the values are to be interpreted.

ABTSTK=ARTIFICIAL BRITTLE STALK. A count of the number of “snapped”plants per plot following machine snapping. A snapped plant has itsstalk completely snapped at a node between the base of the plant and thenode above the ear. Expressed as percent of plants that did not snap.

ALLELE. Any of one or more alternative forms of a genetic sequence. In adiploid cell or organism, the two alleles of a given sequence occupycorresponding loci on a pair of homologous chromosomes.

ANT ROT=ANTHRACNOSE STALK ROT (Colletotrichum graminicola). A 1 to 9visual rating indicating the resistance to Anthracnose Stalk Rot. Ahigher score indicates a higher resistance.

BACKCROSSING. Process in which a breeder crosses a progeny line back toone of the parental genotypes one or more times.

BARPLT=BARREN PLANTS. The percent of plants per plot that was not barren(lack ears).

BREEDING. The genetic manipulation of living organisms.

BRTSTK=BRITTLE STALKS. This is a measure of the stalk breakage near thetime of pollination, and is an indication of whether a hybrid or inbredwould snap or break near the time of flowering under severe winds. Dataare presented as percentage of plants that did not snap.

CLDTST=COLD TEST. The percent of plants that germinate under cold testconditions.

CLN=CORN LETHAL NECROSIS. Synergistic interaction of maize chloroticmottle virus (MCMV) in combination with either maize dwarf mosaic virus(MDMV-A or MDMV-B) or wheat streak mosaic virus (WSMV). A 1 to 9 visualrating indicating the resistance to Corn Lethal Necrosis. A higher scoreindicates a higher resistance.

COMRST=COMMON RUST (Puccinia sorghi). A 1 to 9 visual rating indicatingthe resistance to Common Rust. A higher score indicates a higherresistance.

D/D=DRYDOWN. This represents the relative rate at which a hybrid willreach acceptable harvest moisture compared to other hybrids on a 1-9rating scale. A high score indicates a hybrid that dries relatively fastwhile a low score indicates a hybrid that dries slowly.

DIPERS=DIPLODIA EAR MOLD SCORES (Diplodia maydis and Diplodiamacrospora). A 1 to 9 visual rating indicating the resistance toDiplodia Ear Mold. A higher score indicates a higher resistance.

DIPROT=DIPLODIA STALK ROT SCORE. Score of stalk rot severity due toDiplodia (Diplodia maydis). Expressed as a 1 to 9 score with 9 beinghighly resistant.

DRPEAR=DROPPED EARS. A measure of the number of dropped ears per plotand represents the percentage of plants that did not drop ears prior toharvest.

D/T=DROUGHT TOLERANCE. This represents a 1-9 rating for droughttolerance, and is based on data obtained under stress conditions. A highscore indicates good drought tolerance and a low score indicates poordrought tolerance.

EARHT=EAR HEIGHT. The ear height is a measure from the ground to thehighest placed developed ear node attachment and is measured incentimeters.

EARMLD=General Ear Mold. Visual rating (1-9 score) where a “1” is verysusceptible and a “9” is very resistant. This is based on overall ratingfor ear mold of mature ears without determining the specific moldorganism, and may not be predictive for a specific ear mold.

EARSZ=EAR SIZE. A 1 to 9 visual rating of ear size. The higher therating the larger the ear size.

EBTSTK=EARLY BRITTLE STALK. A count of the number of “snapped” plantsper plot following severe winds when the corn plant is experiencing veryrapid vegetative growth in the V5-V8 stage. Expressed as percent ofplants that did not snap.

ECB1LF=EUROPEAN CORN BORER FIRST GENERATION LEAF FEEDING (Ostrinianubilalis). A 1 to 9 visual rating indicating the resistance topreflowering leaf feeding by first generation European Corn Borer. Ahigher score indicates a higher resistance.

ECB2IT=EUROPEAN CORN BORER SECOND GENERATION INCHES OF TUNNELING(Ostrinia nubilalis). Average inches of tunneling per plant in thestalk.

ECB2SC=EUROPEAN CORN BORER SECOND GENERATION (Ostrinia nubilalis). A 1to 9 visual rating indicating post flowering degree of stalk breakageand other evidence of feeding by European Corn Borer, Second Generation.A higher score indicates a higher resistance.

ECBDPE=EUROPEAN CORN BORER DROPPED EARS (Ostrinia nubilalis). Droppedears due to European Corn Borer. Percentage of plants that did not dropears under second generation corn borer infestation.

EGRWTH=EARLY GROWTH. This is a measure of the relative height and sizeof a corn seedling at the 2-4 leaf stage of growth. This is a visualrating (1 to 9), with 1 being weak or slow growth, 5 being averagegrowth and 9 being strong growth. Taller plants, wider leaves, moregreen mass and darker color constitute higher score.

ELITE INBRED. An inbred that contributed desirable qualities when usedto produce commercial hybrids. An elite inbred may also be used infurther breeding.

ERTLDG=EARLY ROOT LODGING. Early root lodging is the percentage ofplants that do not root lodge prior to or around anthesis; plants thatlean from the vertical axis at an approximately 30° angle or greaterwould be counted as root lodged.

ERTLPN=Early root lodging. An estimate of the percentage of plants thatdo not root lodge prior to or around anthesis; plants that lean from thevertical axis at an approximately 30° angle or greater would beconsidered as root lodged.

ERTLSC=EARLY ROOT LODGING SCORE. Score for severity of plants that leanfrom a vertical axis at an approximate 30-degree angle or greater, whichtypically results from strong winds prior to or around floweringrecorded within 2 weeks of a wind event. Expressed as a 1 to 9 scorewith 9 being no lodging.

ESTCNT=EARLY STAND COUNT. This is a measure of the stand establishmentin the spring and represents the number of plants that emerge on perplot basis for the inbred or hybrid.

EYESPT=Eye Spot (Kabatiella zeae or Aureobasidium zeae). A 1 to 9 visualrating indicating the resistance to Eye Spot. A higher score indicates ahigher resistance.

FUSERS=FUSARIUM EAR ROT SCORE (Fusarium moniliforme or Fusariumsubglutinans). A 1 to 9 visual rating indicating the resistance toFusarium ear rot. A higher score indicates a higher resistance.

GDU=Growing Degree Units. Using the Barger Heat Unit Theory, whichassumes that maize growth occurs in the temperature range 50° F.-86° F.and that temperatures outside this range slow down growth; the maximumdaily heat unit accumulation is 36 and the minimum daily heat unitaccumulation is 0. The seasonal accumulation of GDU is a major factor indetermining maturity zones.

GDUSHD=GDU TO SHED. The number of growing degree units (GDUs) or heatunits required for an inbred line or hybrid to have approximately 50percent of the plants shedding pollen and is measured from the time ofplanting. Growing degree units are calculated by the Barger Method,where the heat units for a 24-hour period are:${GDU} = {\frac{( {{Max}.\quad{temp}.{+ {{Min}.\quad{temp}.}}} )}{2} - 50}$

The highest maximum temperature used is 86° F. and the lowest minimumtemperature used is 50° F. For each inbred or hybrid it takes a certainnumber of GDUs to reach various stages of plant development.

GDUSLK=GDU TO SILK. The number of growing degree units required for aninbred line or hybrid to have approximately 50 percent of the plantswith silk emergence from time of planting. Growing degree units arecalculated by the Barger Method as given in GDU SHD definition.

GENOTYPE. Refers to the genetic constitution of a cell or organism.

GIBERS=GIBBERELLA EAR ROT (PINK MOLD) (Gibberelia zeae). A 1 to 9 visualrating indicating the resistance to Gibberella Ear Rot. A higher scoreindicates a higher resistance.

GIBROT=GIBBERELLA STALK ROT SCORE. Score of stalk rot severity due toGibberella (Gibberella zeae). Expressed as a 1 to 9 score with 9 beinghighly resistant.

GLFSPT=Gray Leaf Spot (Cercospora zeae-maydis). A 1 to 9 visual ratingindicating the resistance to Gray Leaf Spot. A higher score indicates ahigher resistance.

GOSWLT=Goss' Wilt (Corynebacterium nebraskense). A 1 to 9 visual ratingindicating the resistance to Goss' Wilt. A higher score indicates ahigher resistance.

GRNAPP=GRAIN APPEARANCE. This is a 1 to 9 rating for the generalappearance of the shelled grain as it is harvested based on such factorsas the color of harvested grain, any mold on the grain, and any crackedgrain. High scores indicate good grain quality.

H/POP=YIELD AT HIGH DENSITY. Yield ability at relatively high plantdensities on 1-9 relative rating system with a higher number indicatingthe hybrid responds well to high plant densities for yield relative toother hybrids. A 1, 5, and 9 would represent very poor, average, andvery good yield response, respectively, to increased plant density.

HCBLT=HELMINTHOSPORIUM CARBONUM LEAF BLIGHT (Helminthosporium carbonum).A 1 to 9 visual rating indicating the resistance to Helminthosporiuminfection. A higher score indicates a higher resistance.

HD SMT=HEAD SMUT (Sphacelotheca reiliana). This score indicates thepercentage of plants not infected.

HSKCVR=HUSK COVER. A 1 to 9 score based on performance relative to keychecks, with a score of 1 indicating very short husks, tip of ear andkernels showing; 5 is intermediate coverage of the ear under mostconditions, sometimes with thin husk; and a 9 has husks extending andclosed beyond the tip of the ear. Scoring can best be done nearphysiological maturity stage or any time during dry down untilharvested.

INC D/A=GROSS INCOME (DOLLARS PER ACRE). Relative income per acreassuming drying costs of two cents per point above 15.5 percent harvestmoisture and current market price per bushel.

INCOME/ACRE. Income advantage of hybrid to be patented over other hybridon per acre basis.

INC ADV=GROSS INCOME ADVANTAGE. GROSS INCOME advantage of variety #1over variety #2.

KSZDCD=KERNEL SIZE DISCARD. The percent of discard seed; calculated asthe sum of discarded tip kernels and extra large kernels.

LINKAGE. Refers to a phenomenon wherein alleles on the same chromosometend to segregate together more often than expected by chance if theirtransmission was independent.

LINKAGE DISEQUILIBRIUM. Refers to a phenomenon wherein alleles tend toremain together in linkage groups when segregating from parents tooffspring, with a greater frequency than expected from their individualfrequencies.

L/POP=YIELD AT LOW DENSITY. Yield ability at relatively low plantdensities on a 1-9 relative system with a higher number indicating thehybrid responds well to low plant densities for yield relative to otherhybrids. A 1, 5, and 9 would represent very poor, average, and very goodyield response, respectively, to low plant density.

LRTLDG=LATE ROOT LODGING. Late root lodging is the percentage of plantsthat do not root lodge after anthesis through harvest; plants that leanfrom the vertical axis at an approximately 30° angle or greater would becounted as root lodged.

LRTLPN=LATE ROOT LODGING. Late root lodging is an estimate of thepercentage of plants that do not root lodge after anthesis throughharvest; plants that lean from the vertical axis at an approximately 30°angle or greater would be considered as root lodged.

LRTLSC=LATE ROOT LODGING SCORE. Score for severity of plants that leanfrom a vertical axis at an approximate 30-degree angle or greater whichtypically results from strong winds after flowering. Recorded prior toharvest when a root-lodging event has occurred. This lodging results inplants that are leaned or “lodged” over at the base of the plant and donot straighten or “goose-neck” back to a vertical position. Expressed asa 1 to 9 score with 9 being no lodging.

MDMCPX=MAIZE DWARF MOSAIC COMPLEX (MDMV=Maize Dwarf Mosaic Virus andMCDV=Maize Chlorotic Dwarf Virus). A 1 to 9 visual rating indicating theresistance to Maize Dwarf Mosaic Complex. A higher score indicates ahigher resistance.

MST=HARVEST MOISTURE. The moisture is the actual percentage moisture ofthe grain at harvest.

MSTADV=MOISTURE ADVANTAGE. The moisture advantage of variety #1 overvariety #2 as calculated by: MOISTURE of variety #2−MOISTURE of variety#1=MOISTURE ADVANTAGE of variety #1.

NLFBLT=Northern Leaf Blight (Helminthosporium turcicum or Exserohilumturcicum). A 1 to 9 visual rating indicating the resistance to NorthernLeaf Blight. A higher score indicates a higher resistance.

OILT=GRAIN OIL. Absolute value of oil content of the kernel as predictedby Near-infrared Transmittance and expressed as a percent of dry matter.

PEDIGREE DISTANCE. Relationship among generations based on theirancestral links as evidenced in pedigrees. May be measured by thedistance of the pedigree from a given starting point in the ancestry.

PLTHT=PLANT HEIGHT. This is a measure of the height of the plant fromthe ground to the tip of the tassel in centimeters.

POLSC=POLLEN SCORE. A 1 to 9 visual rating indicating the amount ofpollen shed. The higher the score the more pollen shed.

POLWT=POLLEN WEIGHT. This is calculated by dry weight of tasselscollected as shedding commences minus dry weight from similar tasselsharvested after shedding is complete.

POP K/A=PLANT POPULATIONS. Measured as 1000s per acre.

POP ADV=PLANT POPULATION ADVANTAGE. The plant population advantage ofvariety #1 over variety #2 as calculated by PLANT POPULATION of variety#2−PLANT POPULATION of variety #1=PLANT POPULATION ADVANTAGE of variety#1.

PRM=PREDICTED RELATIVE MATURITY. This trait, predicted relativematurity, is based on the harvest moisture of the grain. The relativematurity rating is based on a known set of checks and utilizes standardlinear regression analyses and is also referred to as the ComparativeRelative Maturity Rating System that is similar to the MinnesotaRelative Maturity Rating System.

PRMSHD=A relative measure of the growing degree units (GDU) required toreach 50% pollen shed. Relative values are predicted values from thelinear regression of observed GDU's on relative maturity of commercialchecks.

PROT=GRAIN PROTEIN. Absolute value of protein content of the kernel aspredicted by Near-Infrared Transmittance and expressed as a percent ofdry matter.

RTLDG=ROOT LODGING. Root lodging is the percentage of plants that do notroot lodge; plants that lean from the vertical axis at an approximately30° angle or greater would be counted as root lodged.

RTLADV=ROOT LODGING ADVANTAGE. The root lodging advantage of variety #1over variety #2.

SCTGRN=SCATTER GRAIN. A 1 to 9 visual rating indicating the amount ofscatter grain (lack of pollination or kernel abortion) on the ear. Thehigher the score the less scatter grain.

SDGVGR=SEEDLING VIGOR. This is the visual rating (1 to 9) of the amountof vegetative growth after emergence at the seedling stage(approximately five leaves). A higher score indicates better vigor.

SEL IND=SELECTION INDEX. The selection index gives a single measure ofthe hybrid's worth based on information for up to five traits. A maizebreeder may utilize his or her own set of traits for the selectionindex. One of the traits that is almost always included is yield. Theselection index data presented in the tables represent the mean valueaveraged across testing stations.

SLFBLT=SOUTHERN LEAF BLIGHT (Helminthosporium maydis or Bipolarismaydis). A 1 to 9 visual rating indicating the resistance to SouthernLeaf Blight. A higher score indicates a higher resistance.

SOURST=SOUTHERN RUST (Puccinia polysora). A 1 to 9 visual ratingindicating the resistance to Southern Rust. A higher score indicates ahigher resistance.

STAGRN=STAY GREEN. Stay green is the measure of plant health near thetime of black layer formation (physiological maturity). A high scoreindicates better late-season plant health.

STDADV=STALK STANDING ADVANTAGE. The advantage of variety #1 overvariety #2 for the trait STK CNT.

STKCNT=NUMBER OF PLANTS. This is the final stand or number of plants perplot.

STKLDG=STALK LODGING REGULAR. This is the percentage of plants that didnot stalk lodge (stalk breakage) at regular harvest (when grain moistureis between about 20 and 30%) as measured by either natural lodging orpushing the stalks and determining the percentage of plants that breakbelow the ear.

STKLDL=LATE STALK LODGING. This is the percentage of plants that did notstalk lodge (stalk breakage) at or around late season harvest (whengrain moisture is between about 15 and 18%) as measured by eithernatural lodging or pushing the stalks and determining the percentage ofplants that break below the ear.

STKLDS=STALK LODGING SCORE. A plant is considered as stalk lodged if thestalk is broken or crimped between the ear and the ground. This can becaused by any or a combination of the following: strong winds late inthe season, disease pressure within the stalks, ECB damage orgenetically weak stalks. This trait should be taken just prior to or atharvest. Expressed on a 1 to 9 scale with 9 being no lodging.

STLPCN=STALK LODGING REGULAR. This is an estimate of the percentage ofplants that did not stalk lodge (stalk breakage) at regular harvest(when grain moisture is between about 20 and 30%) as measured by eithernatural lodging or pushing the stalks and determining the percentage ofplants that break below the ear.

STRT=GRAIN STARCH. Absolute value of starch content of the kernel aspredicted by Near-infrared Transmittance and expressed as a percent ofdry matter.

STWWLT=Stewart's Wilt (Erwinia stewartii). A 1 to 9 visual ratingindicating the resistance to Stewart's Wilt. A higher score indicates ahigher resistance.

TASBLS=TASSEL BLAST. A 1 to 9 visual rating was used to measure thedegree of blasting (necrosis due to heat stress) of the tassel at thetime of flowering. A 1 would indicate a very high level of blasting attime of flowering, while a 9 would have no tassel blasting.

TASSZ=TASSEL SIZE. A 1 to 9 visual rating was used to indicate therelative size of the tassel. The higher the rating the larger thetassel.

TAS WT=TASSEL WEIGHT. This is the average weight of a tassel (grams)just prior to pollen shed.

TEXEAR=EAR TEXTURE. A 1 to 9 visual rating was used to indicate therelative hardness (smoothness of crown) of mature grain. A 1 would bevery soft (extreme dent) while a 9 would be very hard (flinty or verysmooth crown).

TILLER=TILLERS. A count of the number of tillers per plot that couldpossibly shed pollen was taken. Data are given as a percentage oftillers: number of tillers per plot divided by number of plants perplot.

TSTWT=TEST WEIGHT (UNADJUSTED). The measure of the weight of the grainin pounds for a given volume (bushel).

TSWADV=TEST WEIGHT ADVANTAGE. The test weight advantage of variety #1over variety #2.

WIN M %=PERCENT MOISTURE WINS.

WIN Y %=PERCENT YIELD WINS.

YIELD BU/A=YIELD (BUSHELS/ACRE). Yield of the grain at harvest inbushels per acre adjusted to 15% moisture.

YLDADV=YIELD ADVANTAGE. The yield advantage of variety #1 over variety#2 as calculated by: YIELD of variety #1−YIELD variety #2=yieldadvantage of variety #1.

YLDSC=YIELD SCORE. A 1 to 9 visual rating was used to give a relativerating for yield based on plot ear piles. The higher the rating thegreater visual yield appearance.

Definitions for Area of Adaptability

When referring to area of adaptability, such term is used to describethe location with the environmental conditions that would be well suitedfor this maize line. Area of adaptability is based on a number offactors, for example: days to maturity, insect resistance, diseaseresistance, and drought resistance. Area of adaptability does notindicate that the maize line will grow in every location within the areaof adaptability or that it will not grow outside the area.

-   Central Corn Belt: Iowa, Illinois, Indiana-   Drylands: non-irrigated areas of North Dakota, South Dakota,    Nebraska, Kansas, Colorado and Oklahoma-   Eastern U.S.: Ohio, Pennsylvania, Delaware, Maryland, Virginia, and    West Virginia-   North central U.S.: Minnesota and Wisconsin-   Northeast: Michigan, New York, Vermont, and Ontario and Quebec    Canada-   Northwest U.S.: North Dakota, South Dakota, Wyoming, Washington,    Oregon, Montana, Utah, and Idaho-   South central U.S.: Missouri, Tennessee, Kentucky, and Arkansas-   Southeast U.S.: North Carolina, South Carolina, Georgia, Florida,    Alabama, Mississippi, and Louisiana-   Southwest U.S.: Texas, Oklahoma, New Mexico, and Arizona-   Western U.S.: Nebraska, Kansas, Colorado, and California-   Maritime Europe: France, Germany, Belgium and Austria

DETAILED DESCRIPTION OF THE INVENTION

Inbred maize lines are typically developed for use in the production ofhybrid maize lines. Inbred maize lines need to be highly homogeneous,substantially homozygous and reproducible to be useful as parents ofcommercial hybrids. There are many analytical methods available todetermine the homozygotic stability and the identity of these inbredlines.

The oldest and most traditional method of analysis is the observation ofphenotypic traits. The data is usually collected in field experimentsover the life of the maize plants to be examined. Phenotypiccharacteristics most often observed are for traits associated with plantmorphology, ear and kernel morphology, insect and disease resistance,maturity, and yield.

In addition to phenotypic observations, the genotype of a plant can alsobe examined. A plant's genotype can be used to identify plants of thesame variety or a related variety. For example, the genotype can be usedto determine the pedigree of a plant. There are many laboratory-basedtechniques available for the analysis, comparison and characterizationof plant genotype; among these are Isozyme Electrophoresis, RestrictionFragment Length Polymorphisms (RFLPs), Randomly Amplified PolymorphicDNAs (RAPDs), Arbitrarily Primed Polymerase Chain Reaction (AP-PCR), DNAAmplification Fingerprinting (DAF), Sequence Characterized AmplifiedRegions (SCARs), Amplified Fragment Length Polymorphisms (AFLPs), SimpleSequence Repeats (SSRs) which are also referred to as Microsatellites,and Single Nucleotide Polymorphisms (SNPs).

Isozyme Electrophoresis and RFLPs as discussed in Lee, M., “Inbred Linesof Maize and Their Molecular Markers,” The Maize Handbook,(Springer-Verlag, New York, Inc. 1994, at 423-432) incorporated hereinby reference, have been widely used to determine genetic composition.Isozyme Electrophoresis has a relatively low number of available markersand a low number of allelic variants among maize inbreds. RFLPs allowmore discrimination because they have a higher degree of allelicvariation in maize and a larger number of markers can be found. Both ofthese methods have been eclipsed by SSRs as discussed in Smith et al.,“An evaluation of the utility of SSR loci as molecular markers in maize(Zea mays L.): comparisons with data from RFLPs and pedigree”,Theoretical and Applied Genetics (1997) vol. 95 at 163-173 and by Pejicet al., “Comparative analysis of genetic similarity among maize inbredsdetected by RFLPs, RAPDs, SSRs, and AFLPs,” Theoretical and AppliedGenetics (1998) at 1248-1255 incorporated herein by reference. SSRtechnology is more efficient and practical to use than RFLPs; moremarker loci can be routinely used and more alleles per marker locus canbe found using SSRs in comparison to RFLPs. Single NucleotidePolymorphisms may also be used to identify the unique geneticcomposition of the invention and progeny lines retaining that uniquegenetic composition. Various molecular marker techniques may be used incombination to enhance overall resolution.

Maize DNA molecular marker linkage maps have been rapidly constructedand widely implemented in genetic studies. One such study is describedin Boppenmaier, et al., “Comparisons among strains of inbreds forRFLPs”, Maize Genetics Cooperative Newsletter, 65:1991, pg. 90, isincorporated herein by reference.

Inbred maize line PH4CV is a yellow, dent maize inbred that is wellsuited to be used as either the female or male in production of thefirst generation F1 maize hybrids. Inbred maize line PH4CV is bestadapted to the Central Corn Belt, Eastern, Southcentral, and Southeastareas of the United States and can be used to produce hybrids withapproximately a 113 maturity based on the Comparative Relative MaturityRating System for harvest moisture of grain. Inbred maize line PH4CVdemonstrates good female yield and good Southern Corn Leaf Blight,Stewarts Bacterial Leaf Blight and Gray Leaf Spot tolerance as an inbredper se. Inbred maize line PH4CV also has above average tolerance toDiplodia, Fusarium, and Gibberella ear rots as an inbred per se. Inhybrid combination, inbred PH4CV demonstrates high grain yield, goodfoliar disease tolerance, and short ear and plant height.

The inbred has shown uniformity and stability within the limits ofenvironmental influence for all the traits as described in the VarietyDescription Information (Table 1) that follows. The inbred has beenself-pollinated and ear-rowed a sufficient number of generations withcareful attention paid to uniformity of plant type to ensure thehomozygosity and phenotypic stability necessary to use in commercialproduction. The line has been increased both by hand and in isolatedfields with continued observation for uniformity. No variant traits havebeen observed or are expected in PH4CV.

Inbred maize line PH4CV, being substantially homozygous, can bereproduced by planting seeds of the line, growing the resulting maizeplants under self-pollinating or sib-pollinating conditions withadequate isolation, and harvesting the resulting seed using techniquesfamiliar to the agricultural arts.

TABLE 1 VARIETY DESCRIPTION INFORMATION VARIETY = PH4CV 1. TYPE: 2 1 =Sweet 2 = Dent 3 = Flint 4 = Flour 5 = Pop 6 = Ornamental 2. MATURITY:DAYS HEAT UNITS 074 1,439.3 From emergence to 50% of plants in silk 0741,451.8 From emergence to 50% of plants in pollen 002 0,067.7 From 10%to 90% pollen shed From 50% silk to harvest at 25% moisture StandardSample 3. PLANT: Deviation  Size  0,221.8 cm Plant Height (to tasseltip) 12.41 45 0,078.3 cm Ear Height (to base of top ear 7.84 45 node)0,015.2 cm Length of Top Ear Internode 1.07 45 0.0 Average Number ofTillers 0.03 9 0.9 Average Number of Ears per Stalk 0.11 9 3.0Anthocyanin of Brace Roots: 1 = Absent 2 = Faint 3 = Moderate 4 = Dark 5= Very Dark Standard Sample 4. LEAF: Deviation  Size  010.5 cm Width ofEar Node Leaf 0.75 45 077.3 cm Length of Ear Node Leaf 3.17 45 07.1Number of leaves above top ear 0.52 45 15.4 Degrees Leaf Angle (measurefrom 4.68 45 2nd leaf above ear at anthesis to stalk above leaf) 03 LeafColor Dark Green (*MC) 7.5GY34 1.2 Leaf Sheath Pubescence (Rate on scalefrom 1 = none to 9 = like peach fuzz) Marginal Waves (Rate on scale from1 = none to 9 = many) Longitudinal Creases (Rate on scale from 1 = noneto 9 = many) Standard Sample 5. TASSEL: Deviation  Size  04.7 Number ofPrimary Lateral 1.63 45 Branches 015.5 Branch Angle from Central Spike6.20 45 50.4 cm Tassel Length (from top leaf collar 3.87 45 to tasseltip) 5.2 Pollen Shed (rate on scale from 0 = male sterile to 9 = heavyshed) 07 Anther Color Yellow (*MC) 10Y96 01 Glume Color Light Green(*MC) 5GY56 1.0 Bar Glumes (Glume Bands): 1 = Absent 2 = Present 17 cmPeduncle Length (cm. from top leaf to basal branches) 6a. EAR (UnhuskedData): 11 Silk Color (3 days after emergence) Pink (*MC) 2.5R58 1 FreshHusk Color (25 days after 50% Light Green (*MC) 5GY56 21 Dry Husk Color(65 days after 50% silking) Buff (*MC) 2.5Y92 3 Position of Ear at DryHusk Stage: 1 = Upright 2 = Horizontal 3 = Pendant 5 Husk Tightness(Rate of Scale from 1 = very loose to 9 = very tight) 2 Husk Extension(at harvest): 1 = Short (ears exposed) 2 = Medium (<8 cm) 3 = Long (8-10cm beyond ear tip) 4 = Very Long (>10 cm) Standard Sample 6b. EAR(Husked Ear Data): Deviation  Size  15 cm Ear Length 0.88 45 42 mm EarDiameter at mid-point 2.01 45 111 gm Ear Weight 23.87 45 16 Number ofKernel Rows 0.50 45 2 Kernel Rows: 1 = Indistinct 2 = Distinct 2 RowAlignment: 1 = Straight 2 = Slightly Curved 3 = Spiral 8 cm Shank Length1.32 45 2 Ear Taper: 1 = Slight 2 = Average 3 = Extreme Standard Sample7. KERNEL (Dried): Deviation  Size  11 mm Kernel Length 1.09 45 7 mmKernel Width 0.71 45 4 mm Kernel Thickness 0.53 45 41 % Round Kernels(Shape Grade) 23.35 9 1 Aleurone Color Pattern: 1 = Homozygous 2 =Segregating 7 Aluerone Color Yellow (*MC) 1.25Y714 7 Hard EndospermColor Yellow (*MC) 10YR612 3 Endosperm Type: Normal Starch 1 = Sweet(Su1) 2 = Extra Sweet (sh2) 3 = Normal Starch 4 = High Amylose Starch 5= Waxy Starch 6 = High Protein 7 = High Lysine 8 = Super Sweet (se) 9 =High Oil 10 = Other     24 gm Weight per 100 Kernels 4.23 9 (unsizedsample) Standard Sample 8. COB: Deviation  Size  23 mm Cob Diameter atmid-point 1.12 45 14 Cob Color Red (*MC) 2.5YR48 9. DISEASE RESISTANCE(Rate from 1 (most susceptible) to 9 (most resistant); leave blank ifnot tested; leave Race or Strain Options blank if polygenic): A. LeafBlights, Wilts, and Local Infection Diseases Anthracnose Leaf Blight(Colletotrichum graminicola) 5 Common Rust (Puccinia Common Smut(Ustilago Eyespot (Kabatiella zeae) Goss's Wilt (Clavibactermichiganense spp. nebraskense) 6 Gray Leaf Spot (Cercospora zeae-maydis)Helminthosporium Leaf Spot (Bipolaris zeicola) Race     5 Northern LeafBlight (Exserohilum turcicum) Race     7 Southern Leaf Blight (Bipolarismaydis) Race     Southern Rust (Puccinia polysora) 6 Stewart's Wilt(Erwinia stewartii) Other (Specify)     B. Systemic Diseases Corn LethalNecrosis (MCMV and MDMV) Head Smut (Sphacelotheca reiliana) MaizeChlorotic Dwarf Virus (MDV) Maize Chlorotic Mottle Virus (MCMV) 4 MaizeDwarf Mosaic Virus (MDMV) Sorghum Downy Mildew of Corn(Peronosclerospora sorghi) Other (Specify)     C. Stalk Rots 3Anthracnose Stalk Rot (Colletotrichum graminicola) Diplodia Stalk Rot(Stenocarpella maydis) Fusarium Stalk Rot (Fusarium moniliforme)Gibberella Stalk Rot (Gibberella zeae) Other (Specify)     D. Ear andKernel Rots Aspergillus Ear and Kernel Rot (Aspergillus flavus) 7Diplodia Ear Rot (Stenocarpella maydis) 6 Fusarium Ear and Kernel Rot(Fusarium moniliforme) 9 Gibberella Ear Rot (Gibberella zeae) Other(Specify)     10. INSECT RESISTANCE (Rate from 1 (most susceptible) to 9(most resistant); (leave blank if not tested): Banks grass Mite(Oligonychus pratensis) Corn Worm (Helicoverpa zea) (sorghi) LeafFeeding (maydis) Silk Feeding  mg larval wt. Ear Damage Corn Leaf Aphid(Rhopalosiphum maidis) Corn Sap Beetle (Carpophilus dimidiatus EuropeanCorn Borer (Ostrinia nubilalis) 9  1st Generation (Typically Whorl LeafFeeding) 4  2nd Generation (Typically Leaf Sheath-Collar  Feeding) Stalk Tunneling  cm tunneled/plant Fall Armyworm (Spodopterafruqiperda)  Leaf Feeding  Silk Feeding  mg larval wt. Maize Weevil(Sitophilus zeamaize Northern Rootworm (Diabrotica barberi) SouthernRootworm (Diabrotica undecimpunctata) Southwestern Corn Borer (Diatreaeagrandiosella)  Leaf Feeding  Stalk Tunneling  cm tunneled/plantTwo-spotted Spider Mite (Tetranychus urticae) Western Rootworm(Diabrotica virgifrea virgifera) Other (Specify)     11. AGRONOMICTRAITS: 3 Staygreen (at 65 days after anthesis)(Rate on a scale from 1 =worst to 0.0 % Dropped Ears (at 65 days after anthesis) % Pre-anthesisBrittle Snapping % Pre-anthesis Root Lodging 80.0 Post-anthesis RootLodging (at 65 days after anthesis) 6,559 Kg/ha Yield (at 12-13% grainmoisture) *MC = Munsell Code (In interpreting the foregoing colordesignations, reference may be made to the Munsell Glossy Book of Color,a standard color reference)Further Embodiments of the Invention

This invention also is directed to methods for producing a maize plantby crossing a first parent maize plant with a second parent maize plantwherein either the first or second parent maize plant is an inbred maizeplant of the line PH4CV. Further, both first and second parent maizeplants can come from the inbred maize line PH4CV. Still further, thisinvention also is directed to methods for producing an inbred maize linePH4CV-derived maize plant by crossing inbred maize line PH4CV with asecond maize plant and growing the progeny seed, and repeating thecrossing and growing steps with the inbred maize line PH4CV-derivedplant from 1 to 2 times, 1 to 3 times 1 to 4 times, or 1 to 5 times.Thus, any such methods using the inbred maize line PH4CV are part ofthis invention: selfing, sibbing, backcrosses, hybrid production,crosses to populations, and the like. All plants produced using inbredmaize line PH4CV as a parent are within the scope of this invention,including plants derived from inbred maize line PH4CV. This includesvarieties essentially derived from variety PH4CV with the term“essentially derived variety” having the meaning ascribed to such termin 7 U.S.C. § 2104(a)(3) of the Plant Variety Protection Act, whichdefinition is hereby incorporated by reference. This also includesprogeny plants and parts thereof with at least one ancestor that isPH4CV, and more specifically, where the pedigree of the progeny includes1, 2, 3, 4, and/or 5 or less cross-pollinations to a maize plant otherthan PH4CV or a plant that has PH4CV as a progenitor. All breeders ofordinary skill in the art maintain pedigree records of their breedingprograms. These pedigree records contain a detailed description of thebreeding process, including a listing of all parental lines used in thebreeding process and information on how such line was used. Thus, abreeder would know if PH4CV were used in the development of a progenyline, and would also know how many crosses to a line other than PH4CV orline with PH4CV as a progenitor were made in the development of anyprogeny line. The inbred maize line may also be used in crosses withother, different, maize inbreds to produce first generation (F₁) maizehybrid seeds and plants with superior characteristics.

Specific methods and products produced using inbred line PH4CV in plantbreeding are encompassed within the scope of the invention listed above.

One such embodiment is a method for developing a PH4CV progeny maizeplant in a maize plant breeding program comprising: obtaining PH4CV orits parts, utilizing said plant or plant parts as a source of breedingmaterial; and selecting a PH4CV progeny plant with molecular markers incommon with PH4CV or morphological and/or physiological characteristicsselected from the characteristics listed in Tables 1 or 2. Breedingsteps that may be used in the maize plant breeding program includepedigree breeding, backcrossing, mutation breeding, and recurrentselection. In conjunction with these steps, techniques such asrestriction fragment polymorphism enhanced selection, genetic markerenhanced selection (for example SSR markers), and the making of doublehaploids may be utilized.

Another such embodiment is the method of crossing inbred maize linePH4CV with another maize plant, such as a different maize inbred line,to form a first generation population of F1 hybrid plants. Thepopulation of first generation F1 hybrid plants produced by this methodis also an embodiment of the invention. This first generation populationof F1 plants will comprise an essentially complete set of the alleles ofinbred line PH4CV. One of ordinary skill in the art can utilize eitherbreeder books or molecular methods to identify a particular F1 hybridplant produced using inbred line PH4CV, and any such individual plant isalso encompassed by this invention. These embodiments also cover use ofthese methods with transgenic or single gene conversions of inbred linePH4CV.

Another such embodiment of this invention is a method of using inbredline PH4CV in breeding that involves the repeated backcrossing to inbredline PH4CV any number of times. Using backcrossing methods, or even thetissue culture and transgenic methods described herein, the single geneconversion methods described herein, or other breeding methods known toone of ordinary skill in the art, one can develop individual plants,plant cells, and populations of plants that retain at least 25%, 30%,35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 79%, 80%, 81%. 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, 99% or 99.5% genetic contribution from inbred line PH4CV. Thepercentage of the genetics retained in the progeny may be measured byeither pedigree analysis or through the use of genetic techniques suchas molecular markers or electrophoresis. In pedigree analysis, onaverage 50% of the starting germplasm would be passed to the progenyline after one cross to another line, 25% after another cross to adifferent line, and so on. Molecular markers could also be used toconfirm and/or determine the pedigree of the progeny line.

One method for producing a line derived from inbred line PH4CV is asfollows. One of ordinary skill in the art would obtain a seed from thecross between inbred line PH4CV and another variety of maize, such as anelite inbred variety. The F1 seed derived from this cross would be grownto form a homogeneous population. The F1 seed would contain essentiallyall of the alleles from variety PH4CV and essentially all of the allelesfrom the other maize variety. The F1 nuclear genome would be made-up of50% variety PH4CV and 50% of the other elite variety. The F1 seed wouldbe grown and allowed to self, thereby forming F2 seed. On average the F2seed would have derived 50% of its alleles from variety PH4CV and 50%from the other maize variety, but many individual plants from thepopulation would have a greater percentage of their alleles derived fromPH4CV (Wang J. and R. Bemardo, 2000, Crop Sci. 40:659-665 and Bemardo,R. and A. L. Kahler, 2001, Theor. Appl. Genet 102:986-992). Themolecular markers of PH4CV could be used to select and retain thoselines with high similarity to PH4CV. The F2 seed would be grown andselection of plants would be made based on visual observation, markersand/or measurement of traits. The traits used for selection may be anyPH4CV trait described in this specification, including the inbred maizePH4CV traits of comparably good yield and comparably good tolerance toleaf blight, leaf spot and ear rot as mentioned herein. Such traits mayalso be the good general or specific combining ability of PH4CV,including its ability to produce hybrids with an approximate 113 CRMmaturity, comparably high yield, comparably good foliar diseasetolerance and comparably short ear and plant height. The PH4CV progenyplants that exhibit one or more of the desired PH4CV traits, such asthose listed above would be selected and each plant would be harvestedseparately. This F3 seed from each plant would be grown in individualrows and allowed to self. Then selected rows or plants from the rowswould be harvested individually. The selections would again be based onvisual observation, markers and/or measurements for desirable traits ofthe plants, such as one or more of the desirable PH4CV traits listedabove. The process of growing and selection would be repeated any numberof times until a PH4CV progeny inbred plant is obtained. The PH4CVprogeny inbred plant would contain desirable traits derived from inbredplant PH4CV, some of which may not have been expressed by the othermaize variety to which inbred line PH4CV was crossed and some of whichmay have been expressed by both maize varieties but now would be at alevel equal to or greater than the level expressed in inbred varietyPH4CV. However, in each case the resulting progeny line would benefitfrom the efforts of the inventor(s), and would not have existed but forthe inventor(s) work in creating PH4CV. The PH4CV progeny inbred plantswould have, on average, 50% of their nuclear genes derived from inbredline PH4CV, but many individual plants from the population would have agreater percentage of their alleles derived from PH4CV. This breedingcycle, of crossing and selfing, and optional selection, may be repeatedto produce another population of PH4CV progeny maize plants with, onaverage, 25% of their nuclear genes derived from inbred line PH4CV, but,again, many individual plants from the population would have a greaterpercentage of their alleles derived from PH4CV. Another embodiment ofthe invention is a PH4CV progeny plant that has received the desirablePH4CV traits listed above through the use of PH4CV, which traits werenot exhibited by other plants used in the breeding process.

The previous example can be modified in numerous ways, for instanceselection may or may not occur at every selfing generation, selectionmay occur before or after the actual self-pollination process occurs, orindividual selections may be made by harvesting individual ears, plants,rows or plots at any point during the breeding process described. Inaddition, double haploid breeding methods may be used at any step in theprocess. The population of plants produced at each and any cycle ofbreeding is also an embodiment of the invention, and on average eachsuch population would predictably consist of plants containingapproximately 50% of its genes from inbred line PH4CV in the firstbreeding cycle, 25% of its genes from inbred line PH4CV in the secondbreeding cycle, 12.5% of its genes from inbred line PH4CV in the thirdbreeding cycle and so on. However, in each case the use of PH4CVprovides a substantial benefit. The linkage groups of PH4CV would beretained in the progeny lines, and since current estimates of the maizegenome size is about 50,000-80,000 genes (Xiaowu, Gai et al., NucleicAcids Research, 2000, Vol. 28, No. 1, 94-96), in addition to non-codingDNA that impacts gene expression, it provides a significant advantage touse PH4CV as starting material to produce a line that retains desiredgenetics or traits of PH4CV.

Another embodiment of this invention is the method of obtaining asubstantially homozygous PH4CV progeny plant by obtaining a seed fromthe cross of PH4CV and another maize plant and applying double haploidmethods to the F1 seed or F1 plant or to any successive filialgeneration. Such methods decrease the number of generations required toproduce an inbred with similar genetics or characteristics to PH4CV. SeeBernardo, R. and Kahler, A. L., Theor. Appl. Genet. 102:986-992, 2001.

A further embodiment of the invention is a single gene conversion ofPH4CV. A single gene conversion occurs when DNA sequences are introducedthrough traditional (non-transformation) breeding techniques, such asbackcrossing (Hallauer et al, 1988). DNA sequences, whether naturallyoccurring or transgenes, may be introduced using these traditionalbreeding techniques. The term single gene conversion is also referred toin the art as a single locus conversion. Reference is made to US2002/0062506A1 for a detailed discussion of single locus conversions andtraits that may be incorporated into PH4CV through single geneconversion. Desired traits transferred through this process include, butare not limited to, waxy starch, nutritional enhancements, industrialenhancements, disease resistance, insect resistance, herbicideresistance and yield enhancements. The trait of interest is transferredfrom the donor parent to the recurrent parent, in this case, the maizeplant disclosed herein. Single gene traits may result from either thetransfer of a dominant allele or a recessive allele. Selection ofprogeny containing the trait of interest is accomplished by directselection for a trait associated with a dominant allele. Selection ofprogeny for a trait that is transferred via a recessive allele, such asthe waxy starch characteristic, requires growing and selfing the firstbackcross generation to determine which plants carry the recessivealleles. Recessive traits may require additional progeny testing insuccessive backcross generations to determine the presence of the geneof interest. Along with selection for the trait of interest, progeny areselected for the phenotype of the recurrent parent. It should beunderstood that occasionally additional polynucleotide sequences orgenes are transferred along with the single gene conversion trait ofinterest. A progeny comprising at least 98%, 99%, 99.5% and 99.9% of thegenes from the recurrent parent, the maize line disclosed herein, pluscontaining the single gene conversion trait or traits of interest, isconsidered to be a single gene conversion of inbred line PH4CV.

It should be understood that the inbred could, through routinemanipulation by detasseling, cytoplasmic genes, nuclear genes, or otherfactors, be produced in a male-sterile form. Such embodiments are alsowithin the scope of the present claims. The term manipulated to be malesterile refers to the use of any available techniques to produce a malesterile version of maize line PH4CV. The male sterility may be eitherpartial or complete male sterility.

This invention is also directed to the use of PH4CV in tissue culture.As used herein, the term plant includes plant protoplasts, plant celltissue cultures from which maize plants can be regenerated, plant calli,plant clumps, and plant cells that are intact in plants or parts ofplants, such as embryos, pollen, ovules, seeds, flowers, kernels, ears,cobs, leaves, husks, stalks, roots, root tips, anthers, silk and thelike. As used herein, phrases such as “growing the seed” or “grown fromthe seed” include embryo rescue, isolation of cells from seed for use intissue culture, as well as traditional growing methods.

Duncan, Williams, Zehr, and Widholm, Planta (1985)165:322-332 reflectsthat 97% of the plants cultured that produced callus were capable ofplant regeneration. Subsequent experiments with both inbreds and hybridsproduced 91% regenerable callus that produced plants. In a further studyin 1988, Songstad, Duncan & Widholm in Plant Cell Reports (1988),7:262-265 reports several media additions that enhance regenerability ofcallus of two inbred lines. Other published reports also indicated that“nontraditional” tissues are capable of producing somatic embryogenesisand plant regeneration. K. P. Rao, et al., Maize Genetics CooperationNewsletter, 60:64-65 (1986), refers to somatic embryogenesis from glumecallus cultures and B. V. Conger, et al., Plant Cell Reports, 6:345-347(1987) indicates somatic embryogenesis from the tissue cultures of maizeleaf segments. Thus, it is clear from the literature that the state ofthe art is such that these methods of obtaining plants are, and were,“conventional” in the sense that they are routinely used and have a veryhigh rate of success.

Tissue culture of maize, including tassel/anther culture, is describedin U.S. 2002/0062506A1 and European Patent Application, publication160,390, each of which is incorporated herein by reference. Maize tissueculture procedures are also described in Green and Rhodes, “PlantRegeneration in Tissue Culture of Maize,” Maize for Biological Research(Plant Molecular Biology Association, Charlottesville, Va. 1982, at367-372) and in Duncan, et al., “The Production of Callus Capable ofPlant Regeneration from Immature Embryos of Numerous Zea MaysGenotypes,” 165 Planta 322-332 (1985). Thus, another aspect of thisinvention is to provide cells, which upon growth and differentiationproduce maize plants having the genotype and/or physiological andmorphological characteristics of inbred line PH4CV.

The utility of inbred maize line PH4CV also extends to crosses withother species. Commonly, suitable species will be of the familyGraminaceae, and especially of the genera Zea, Tripsacum, Coix,Schlerachne, Polytoca, Chionachne, and Trilobachne, of the tribeMaydeae. Potentially suitable for crosses with PH4CV may be the variousvarieties of grain sorghum, Sorghum bicolor (L.) Moench.

The advent of new molecular biological techniques has allowed theisolation and characterization of genetic elements with specificfunctions, such as encoding specific protein products. Scientists in thefield of plant biology developed a strong interest in engineering thegenome of plants to contain and express foreign genetic elements, oradditional, or modifed versions of native or endogenous genetic elementsin order to alter the traits of a plant in a specific manner. Any DNAsequences, whether from a different species or from the same speciesthat are inserted into the genome using transformation are referred toherein collectively as “transgenes”. Over the last fifteen to twentyyears several methods for producing transgenic plants have beendeveloped, and the present invention, in particular embodiments, alsorelates to transformed versions of the claimed inbred maize line PH4CV.

Numerous methods for plant transformation have been developed, includingbiological and physical, plant transformation protocols. See, forexample, Miki et al., “Procedures for Introducing Foreign DNA intoPlants” in Methods in Plant Molecular Biology and Biotechnology, Glick,B. R. and Thompson, J. E. Eds. (CRC Press, Inc., Boca Raton, 1993) pages67-88 and Armstrong, “The First Decade of Maize Transformation: A Reviewand Future Perspective” (Maydica 44:101-109, 1999). In addition,expression vectors and in vitro culture methods for plant cell or tissuetransformation and regeneration of plants are available. See, forexample, Gruber et al., “Vectors for Plant Transformation” in Methods inPlant Molecular Biology and Biotechnology, Glick, B. R. and Thompson, J.E. Eds. (CRC Press, Inc., Boca Raton, 1993) pages 89-119. See U.S. Pat.No. 6,118,055, which is herein incorporated by reference.

The most prevalent types of plant transformation involve theconstruction of an expression vector. Such a vector comprises a DNAsequence that contains a gene under the control of or operatively linkedto a regulatory element, for example a promoter. The vector may containone or more genes and one or more regulatory elements.

A genetic trait which has been engineered into a particular maize plantusing transformation techniques, could be moved into another line usingtraditional breeding techniques that are well known in the plantbreeding arts. For example, a backcrossing approach could be used tomove a transgene from a transformed maize plant to an elite inbred lineand the resulting progeny would comprise a transgene. Also, if an inbredline was used for the transformation then the transgenic plants could becrossed to a different inbred in order to produce a transgenic hybridmaize plant. As used herein, “crossing” can refer to a simple X by Ycross, or the process of backcrossing, depending on the context.

Various genetic elements can be introduced into the plant genome usingtransformation. These elements include but are not limited to genes;coding sequences; inducible, constitutive, and tissue specificpromoters; enhancing sequences; and signal and targeting sequences. SeeU.S. Pat. No. 6,118,055, which is herein incorporated by reference.

With transgenic plants according to the present invention, a foreignprotein can be produced in commercial quantities. Thus, techniques forthe selection and propagation of transformed plants, which are wellunderstood in the art, yield a plurality of transgenic plants, which areharvested in a conventional manner, and a foreign protein then can beextracted from a tissue of interest or from total biomass. Proteinextraction from plant biomass can be accomplished by known methods,which are discussed, for example, by Heney and Orr, Anal. Biochem. 114:92-6 (1981).

According to a preferred embodiment, the transgenic plant provided forcommercial production of foreign protein is maize. In another preferredembodiment, the biomass of interest is seed. A genetic map can begenerated, primarily via conventional Restriction Fragment LengthPolymorphisms (RFLP), Polymerase Chain Reaction (PCR) analysis, andSimple Sequence Repeats (SSR) and Single Nucleotide Polymorphisms (SNP),which identify the approximate chromosomal location of the integratedDNA molecule. For exemplary methodologies in this regard, see Glick andThompson, METHODS IN PLANT MOLECULAR BIOLOGY AND BIOTECHNOLOGY 269-284(CRC Press, Boca Raton, 1993).

Wang et al. discuss “Large Scale Identification, Mapping and Genotypingof Single-Nucleotide Polymorphorsms in the Human Genome”, Science,280:1077-1082, 1998, and similar capabilities will soon be available forthe corn genome. Map information concerning chromosomal location isuseful for proprietary protection of a subject transgenic plant. Ifunauthorized propagation is undertaken and crosses made with othergermplasm, the map of the integration region can be compared to similarmaps for suspect plants, to determine if the latter have a commonparentage with the subject plant. Map comparisons would involvehybridizations, RFLP, PCR, SSR and sequencing, all of which areconventional techniques. SNP's may also be used alone or in combinationwith other techniques.

Likewise, by means of the present invention, plants can be geneticallyengineered to express various phenotypes of agronomic interest. Throughthe transformation of maize the expression of genes can be modulated toenhance disease resistance, insect resistance, herbicide resistance,agronomic traits as well as grain quality traits. Transformation canalso be used to insert DNA sequences which control or help controlmale-sterility. DNA sequences native to maize as well as non-native DNAsequences can be transformed into maize and used to modulate levels ofnative or non-native proteins. Anti-sense technology, various promoters,targeting sequences, enhancing sequences, and other DNA sequences can beinserted into the maize genome for the purpose of modulating theexpression of proteins. Exemplary transgenes implicated in this regardinclude, but are not limited to, those categorized below.

1. Transgenes that Confer Resistance to Pests or Disease and thatEncode:

(A) Plant disease resistance genes. Plant defenses are often activatedby specific interaction between the product of a disease resistance gene(R) in the plant and the product of a corresponding avirulence (Avr)gene in the pathogen. A plant variety can be transformed with clonedresistance gene to engineer plants that are resistant to specificpathogen strains. See, for example Jones et al., Science 266: 789 (1994)(cloning of the tomato Cf-9 gene for resistance to Cladosporium fulvum);Martin et al., Science 262: 1432 (1993) (tomato Pto gene for resistanceto Pseudomonas syringae pv. tomato encodes a protein kinase); Mindrinoset al., Cell 78: 1089 (1994) (Arabidopsis RSP2 gene for resistance toPseudomonas syringae).

(B) A Bacillus thuringiensis protein, a derivative thereof or asynthetic polypeptide modeled thereon. See, for example, Geiser et al.,Gene 48: 109 (1986), who disclose the cloning and nucleotide sequence ofa Bt δ-endotoxin gene. Moreover, DNA molecules encoding δ-endotoxingenes can be purchased from American Type Culture Collection (Rockville,Md.), for example, under ATCC Accession Nos. 40098, 67136, 31995 and31998. Other examples of Bacillus thuringiensis transgenes beinggenetically engineered are given in the following patents and hereby areincorporated by reference: U.S. Pat. Nos. 5,188,960; 5,689,052;5,880,275; and WO 97/40162.

(C) A lectin. See, for example, the disclosure by Van Damme et al.,Plant Molec. Biol. 24: 25 (1994), who disclose the nucleotide sequencesof several Clivia miniata mannose-binding lectin genes.

(D) A vitamin-binding protein such as avidin. See PCT applicationUS93/06487 the contents of which are hereby incorporated by reference.The application teaches the use of avidin and avidin homologues aslarvicides against insect pests.

(E) An enzyme inhibitor, for example, a protease inhibitor or an amylaseinhibitor. See, for example, Abe et al., J. Biol. Chem. 262: 16793(1987) (nucleotide sequence of rice cysteine proteinase inhibitor), Huubet al., Plant Molec. Biol. 21: 985 (1993) (nucleotide sequence of cDNAencoding tobacco proteinase inhibitor I), and Sumitani et al., Biosci.Biotech. Biochem. 57: 1243 (1993) (nucleotide sequence of Streptomycesnitrosporeus α-amylase inhibitor) and U.S. Pat. No. 5,494,813.

(F) An insect-specific hormone or pheromone such as an ecdysteroid andjuvenile hormone, a variant thereof, a mimetic based thereon, or anantagonist or agonist thereof. See, for example, the disclosure byHammock et al., Nature 344: 458 (1990), of baculovirus expression ofcloned juvenile hormone esterase, an inactivator of juvenile hormone.

(G) An insect-specific peptide or neuropeptide, which, upon expression,disrupts the physiology of the affected pest. For example, see thedisclosures of Regan, J. Biol. Chem. 269: 9 (1994) (expression cloningyields DNA coding for insect diuretic hormone receptor), and Pratt etal., Biochem. Biophys. Res. Comm. 163: 1243 (1989) (an allostatin isidentified in Diploptera puntata). See also U.S. Pat. No. 5,266,317 toTomalski et al., who disclose genes encoding insect-specific, paralyticneurotoxins.

(H) An insect-specific venom produced in nature by a snake, a wasp, etc.For example, see Pang et al., Gene 116: 165 (1992), for disclosure ofheterologous expression in plants of a gene coding for a scorpioninsectotoxic peptide.

(I) An enzyme responsible for an hyperaccumulation of a monterpene, asesquiterpene, a steroid, hydroxamic acid, a phenylpropanoid derivativeor another non-protein molecule with insecticidal activity.

(J) An enzyme involved in the modification, including thepost-translational modification, of a biologically active molecule; forexample, a glycolytic enzyme, a proteolytic enzyme, a lipolytic enzyme,a nuclease, a cyclase, a transaminase, an esterase, a hydrolase, aphosphatase, a kinase, a phosphorylase, a polymerase, an elastase, achitinase and a glucanase, whether natural or synthetic. See PCTapplication WO 93/02197 in the name of Scott et al., which discloses thenucleotide sequence of a callase gene. DNA molecules which containchitinase-encoding sequences can be obtained, for example, from the ATCCunder Accession Nos. 39637 and 67152. See also Kramer et al., InsectBiochem. Molec. Biol. 23: 691 (1993), who teach the nucleotide sequenceof a cDNA encoding tobacco hookworm chitinase, and Kawalleck et al.,Plant Molec. Biol. 21: 673 (1993), who provide the nucleotide sequenceof the parsley ubi4-2 polyubiquitin gene.

(K) A molecule that stimulates signal transduction. For example, see thedisclosure by Botella et al., Plant Molec. Biol. 24: 757 (1994), ofnucleotide sequences for mung bean calmodulin cDNA clones, and Griess etal., Plant Physiol. 104: 1467 (1994), who provide the nucleotidesequence of a maize calmodulin cDNA clone.

(L) A hydrophobic moment peptide. See PCT application WO95/16776(disclosure of peptide derivatives of Tachyplesin which inhibit fungalplant pathogens) and PCT application WO95/18855 (teaches syntheticantimicrobial peptides that confer disease resistance), the respectivecontents of which are hereby incorporated by reference.

(M) A membrane permease, a channel former or a channel blocker. Forexample, see the disclosure by Jaynes et al., Plant Sci. 89: 43 (1993),of heterologous expression of a cecropin-βlytic peptide analog to rendertransgenic tobacco plants resistant to Pseudomonas solanacearum.

(N) A viral-invasive protein or a complex toxin derived therefrom. Forexample, the accumulation of viral coat proteins in transformed plantcells imparts resistance to viral infection and/or disease developmenteffected by the virus from which the coat protein gene is derived, aswell as by related viruses. See Beachy et al., Ann. Rev. Phytopathol.28: 451 (1990). Coat protein-mediated resistance has been conferred upontransformed plants against alfalfa mosaic virus, cucumber mosaic virus,tobacco streak virus, potato virus X, potato virus Y, tobacco etchvirus, tobacco rattle virus and tobacco mosaic virus. Id.

(O) An insect-specific antibody or an immunotoxin derived therefrom.Thus, an antibody targeted to a critical metabolic function in theinsect gut would inactivate an affected enzyme, killing the insect. Cf.Taylor et al., Abstract #497, SEVENTH INT'L SYMPOSIUM ON MOLECULARPLANT-MICROBE INTERACTIONS (Edinburgh, Scotland, 1994) (enzymaticinactivation in transgenic tobacco via production of single-chainantibody fragments).

(P) A virus-specific antibody. See, for example, Tavladoraki et al.,Nature 366: 469 (1993), who show that transgenic plants expressingrecombinant antibody genes are protected from virus attack.

(Q) A developmental-arrestive protein produced in nature by a pathogenor a parasite. Thus, fungal endo α-1,4-D-polygalacturonases facilitatefungal colonization and plant nutrient release by solubilizing plantcell wall homo-α-1,4-D-galacturonase. See Lamb et al., Bio/Technology10: 1436 (1992). The cloning and characterization of a gene whichencodesa bean endopolygalacturonase-inhibiting protein is described by Toubartet al., Plant J. 2: 367 (1992).

(R) A developmental-arrestive protein produced in nature by a plant. Forexample, Logemann et al., Bio/Technology 10: 305 (1992), have shown thattransgenic plants expressing the barley ribosome-inactivating gene havean increased resistance to fungal disease.

(S) Genes involved in the Systemic Acquired Resistance (SAR) Responseand/or the pathogenesis related genes. Briggs, S., Current Biology, 5(2)(1995).

(T) Antifungal genes (Cornelissen and Melchers, Pl. Physiol.101:709-712, (1993) and Parijs et al., Planta 183:258-264, (1991) andBushnell et al., Can. J. of Plant Path. 20(2):137-149 (1998).

2. Transgenes that Confer Resistance to a Herbicide, for Example:

(A)A herbicide that inhibits the growing point or meristem, such as animidazolinone or a sulfonylurea. Exemplary genes in this category codefor mutant ALS and AHAS enzyme as described, for example, by Lee et al.,EMBO J. 7: 1241 (1988), and Miki et al., Theor. Appl. Genet. 80: 449(1990), respectively. See also, U.S. Pat. Nos. 5,605,011; 5,013,659;5,141,870; 5,767,361; 5,731,180; 5,304,732; 4,761,373; 5,331,107;5,928,937; and 5,378,824; and international publication WO 96/33270,which are incorporated herein by reference in their entireties for allpurposes.

(B) Glyphosate (resistance imparted by mutant5-enolpyruvl-3-phosphikimate synthase (EPSP), and aroA genes,respectively) and other phosphono compounds such as glufosinate(phosphinothricin acetyl transferase (PAT) and Streptomyceshygroscopicus phosphinothricin acetyl transferase (bar) genes), andpyridinoxy or phenoxy proprionic acids and cycloshexones (ACCaseinhibitor-encoding genes). See, for example, U.S. Pat. No. 4,940,835 toShah et al., which discloses the nucleotide sequence of a form of EPSPS,which can confer glyphosate resistance. U.S. Pat. No. 5,627,061 to Barryet al. also describes genes encoding EPSPS enzymes. See also U.S. Pat.Nos. 6,248,876 B1; 6,040,497; 5,804,425; 5,633,435; 5,145,783;4,971,908; 5,312,910; 5,188,642; 4,940,835; 5,866,775; 6,225,114 B1;6,130,366; 5,310,667; 4,535,060; 4,769,061; 5,633,448; 5,510,471; Re.36,449; RE 37,287 E; and U.S. Pat. No. 5,491,288; and internationalpublications WO 97/04103; WO 97/04114; WO 00/66746; WO 01/66704; WO00/66747 and WO 00/66748, which are incorporated herein by reference intheir entirety. Glyphosate resistance is also imparted to plants thatexpress a gene that encodes a glyphosate oxidoreductase enzyme asdescribed more fully in U.S. Pat. Nos. 5,776,760 and 5,463,175, whichare incorporated herein by reference in their entirety. In additionglyphosate resistance can be imparted to plants by the over expressionof genes encoding glyphosate N-acetyltransferase. See, for example, U.S.Application Ser. Nos. 60/244,385; 60/377,175 and 60/377,719.

A DNA molecule encoding a mutant aroA gene can be obtained under ATCCaccession No. 39256, and the nucleotide sequence of the mutant gene isdisclosed in U.S. Pat. No. 4,769,061 to Comai. European patentapplication No. 0 333 033 to Kumada et al. and U.S. Pat. No. 4,975,374to Goodman et al. disclose nucleotide sequences of glutamine synthetasegenes which confer resistance to herbicides such as L-phosphinothricin.The nucleotide sequence of a phosphinothricin-acetyl-transferase gene isprovided in European patent No. 0 242 246 and 0 242 236 to Leemans etal. De Greef et al., Bio/Technology 7: 61 (1989), describe theproduction of transgenic plants that express chimeric bar genes codingfor phosphinothricin acetyl transferase activity. See also, U.S. Pat.Nos. 5,969,213; 5,489,520; 5,550,318; 5,874,265; 5,919,675; 5,561,236;5,648,477; 5,646,024; 6,177,616 B1; and 5,879,903, which areincorporated herein by reference in their entirety. Exemplary of genesconferring resistance to phenoxy proprionic acids and cycloshexones,such as sethoxydim and haloxyfop, are the Acc1-S1, Acc1-S2 and Acc1-S3genes described by Marshall et al., Theor. Appl. Genet. 83: 435 (1992).

(C) A herbicide that inhibits photosynthesis, such as a triazine (psbAand gs+genes) and a benzonitrile (nitrilase gene). Przibilla et al.,Plant Cell 3: 169 (1991), describe the transformation of Chlamydomonaswith plasmids encoding mutant psbA genes. Nucleotide sequences fornitrilase genes are disclosed in U.S. Pat. No. 4,810,648 to Stalker, andDNA molecules containing these genes are available under ATCC AccessionNos. 53435, 67441 and 67442. Cloning and expression of DNA coding for aglutathione S-transferase is described by Hayes et al, Biochem. J. 285:173 (1992).

(D) Acetohydroxy acid synthase, which has been found to make plants thatexpress this enzyme resistant to multiple types of herbicides, has beenintroduced into a variety of plants (see, e.g., Hattori et al. (1995)Mol Gen Genet 246:419). Other genes that confer tolerance to herbicidesinclude: a gene encoding a chimeric protein of rat cytochrome P4507A1and yeast NADPH-cytochrome P450 oxidoreductase (Shiota et al. (1994)Plant PhysiolPlant Physiol 106:17), genes for glutathione reductase andsuperoxide dismutase (Aono et al. (1995) Plant Cell Physiol 36:1687, andgenes for various phosphotransferases (Datta et al. (1992) Plant MolBiol 20:619).

(E) Protoporphyrinogen oxidase (protox) is necessary for the productionof chlorophyll, which is necessary for all plant survival. The protoxenzyme serves as the target for a variety of herbicidal compounds. Theseherbicides also inhibit growth of all the different species of plantspresent, causing their total destruction. The development of plantscontaining altered protox activity which are resistant to theseherbicides are described in U.S. Pat. Nos. 6,288,306 B1; 6,282,837 B1;and 5,767,373; and international publication WO 01/12825, which areincorporated herein by reference in their entireties of all purposes.

3. Transgenes that Confer or Contribute to a Value-Added Trait, Such as:

(A) Modified fatty acid metabolism, for example, by transforming a plantwith an antisense gene of steary-ACP desaturase to increase stearic acidcontent of the plant. See Knultzon et al., Proc. Nati. Acad. Sci. USA89: 2624 (1992).

(B) Decreased phytate content

-   -   (1) Introduction of a phytase-encoding gene would enhance        breakdown of phytate, adding more free phosphate to the        transformed plant. For example, see Van Hartingsveldt et al.,        Gene 127: 87 (1993), for a disclosure of the nucleotide sequence        of an Aspergillus niger phytase gene.    -   (2) A gene could be introduced that reduces phytate content. In        maize, this, for example, could be accomplished, by cloning and        then re-introducing DNA associated with the single allele which        is responsible for maize mutants characterized by low levels of        phytic acid. See Raboy et al., Maydica 35: 383 (1990).

(C) Modified carbohydrate composition effected, for example, bytransforming plants with a gene coding for an enzyme that alters thebranching pattern of starch. See Shiroza et al., J. Bacteriol. 170: 810(1988) (nucleotide sequence of Streptococcus mutans fructosyltransferasegene), Steinmetz et al., Mol. Gen. Genet. 200: 220 (1985) (nucleotidesequence of Bacillus subtilis levansucrase gene), Pen et al.,Bio/Technology 10: 292 (1992) (production of transgenic plants thatexpress Bacillus licheniformis α-amylase), Elliot et al., Plant Molec.Biol. 21: 515 (1993) (nucleotide sequences of tomato invertase genes),Sφgaard et al., J. Biol. Chem. 268: 22480 (1993) (site-directedmutagenesis of barley α-amylase gene), and Fisher et al., Plant Physiol.102: 1045 (1993) (maize endosperm starch branching enzyme II).

(D) Elevated oleic acid via FAD-2 gene modification and/or decreasedlinolenic acid via FAD-3 gene modification (see U.S. Pat. Nos.6,063,947; 6,323,392; and WO 93/11245).

4. Genes that Control Male-Sterility

(A) Introduction of a deacetylase gene under the control of atapetum-specific promoter and with the application of the chemicalN-Ac-PPT (WO 01/29237).

(B) Introduction of various stamen-specific promoters (WO 92/13956, WO92/13957).

(C) Introduction of the bamase and the barstar gene (Paul et al. PlantMol. Biol. 19:611-622, 1992).

INDUSTRIAL APPLICABILITY

Maize is used as human food, livestock feed, and as raw material inindustry. The food uses of maize, in addition to human consumption ofmaize kernels, include both products of dry- and wet-milling industries.The principal products of maize dry milling are grits, meal and flour.The maize wet-milling industry can provide maize starch, maize syrups,and dextrose for food use. Maize oil is recovered from maize germ, whichis a by-product of both dry- and wet-milling industries.

Maize, including both grain and non-grain portions of the plant, is alsoused extensively as livestock feed, primarily for beef cattle, dairycattle, hogs, and poultry.

Industrial uses of maize include production of ethanol, maize starch inthe wet-milling industry and maize flour in the dry-milling industry.The industrial applications of maize starch and flour are based onfunctional properties, such as viscosity, film formation, adhesiveproperties, and ability to suspend particles. The maize starch and flourhave application in the paper and textile industries. Other industrialuses include applications in adhesives, building materials, foundrybinders, laundry starches, explosives, oil-well muds, and other miningapplications.

Plant parts other than the grain of maize are also used in industry: forexample, stalks and husks are made into paper and wallboard and cobs areused for fuel and to make charcoal.

The seed of inbred maize line PH4CV, the plant produced from the inbredseed, the hybrid maize plant produced from the crossing of the inbred,hybrid seed, and various parts of the hybrid maize plant and transgenicversions of the foregoing, can be utilized for human food, livestockfeed, and as a raw material in industry.

PERFORMANCE EXAMPLES OF PH4CV

In the examples that follow, data from traits and characteristics ofinbred maize line PH4CV per se and in a hybrid are given and compared toother maize inbred lines and hybrids.

Inbred Comparisons

The results in Table 2A compare inbred PH4CV to inbred PHBE2. Theresults show inbred PH4CV produced significantly higher yield. InbredPH4CV demonstrated significantly higher cold test scores than PHBE2.Inbred PH4CV also demonstrated significantly better Gray Leaf Spottolerance scores and Southern Leaf Blight tolerance scores than PHBE2.

The results in Table 2B compare inbred PH4CV to inbred PHR03. Theresults show inbred PH4CV produced significantly higher yield andsignificantly lower harvest moisture of grain. Inbred PH4CV also showedsignificantly better early stand count scores, Gray Leaf Spot tolerancescores, and Southern Leaf Blight tolerance scores than PHR03.

The results in Table 2C compare inbred PH4CV to inbred PH1BC. Theresults show inbred PH4CV had significantly shorter plant height. InbredPH4CV demonstrated significantly better scores for Southern Leaf Blighttolerance and Maize Dwarf Mosaic Complex resistance than PH1BC.

The results in Table 2D compare PH4CV to inbred PH2EJ. The results showinbred PH4CV produced significantly higher yield. Inbred PH4CV alsodemonstrated significantly better scores than PH2EJ for early growth,stay green, Gray Leaf Spot tolerance, and Southern Leaf Blighttolerance.

Hybrid Comparisons

The results in Table 3A compare a hybrid in which inbred PH4CV is aparent and a second hybrid, 33J56. The results show that the hybridcontaining PH4CV produced significantly higher yield. The hybridcontaining PH4CV grew to a significantly shorter plant height and hadsignificantly lower placement of the ear than 33J56. The hybridcontaining PH4CV also demonstrated significantly better scores than33J56 for stay green and Fusarium ear rot tolerance. The hybridcontaining PH4CV also had significantly less stalk lodging than 33J56.

The results in Table 3B compare a hybrid in which inbred PH4CV is aparent and a second hybrid, 3335. The results show that the hybridcontaining PH4CV produced significantly higher yield.

The results in Table 3C compare a hybrid in which inbred PH4CV is aparent and a second hybrid, 3245. The results show that the hybridcontaining PH4CV produced significantly higher yield. The hybridcontaining PH4CV grew to a significantly shorter plant than hybrid 3245.The hybrid containing PH4CV also had significantly better stay greenscores than 3245.

The results in Table 3D compare a hybrid in which inbred PH4CV is aparent and a second hybrid, 32H58. The results show that the hybridcontaining PH4CV produced significantly higher yield. The hybridcontaining PH4CV also demonstrated significantly better scores than32H58 for stay green, Southern Leaf Blight tolerance, husk cover andstalk lodging.

The results in Table 3E compare a hybrid in which inbred PH4CV is aparent and a second hybrid, 31G98. The results show that the hybridcontaining PH4CV grew to a significantly shorter plant height and hadsignificantly lower placement of the ear than hybrid 31G98. Thecomparison also showed that the PH4CV-containing hybrid hadsignificantly less stalk lodging and brittle snap than 31G98. The hybridcontaining PH4CV also demonstrated significantly better scores thanhybrid 31G98 for husk cover, Southern Leaf Blight tolerance and Fusariumear rot tolerance.

TABLE 2A PAIRED INBRED CQMPARISON REPORT Variety #1: PH4CV Variety #2:PHBE2 YIELD YIELD MST TSTWT EGRWTH ESTCNT BU/A 56# BU/A 56# PCT LB/BUSCORE COUNT Stat ABS % MN ABS ABS ABS ABS Mean 1 94.5 126.9 19.1 58.25.5 20.4 Mean 2 81.8 109.6 16.7 60.0 5.3 18.9 Locs 19 19 20 5 30 21 Reps19 19 20 5 30 21 Diff 12.7 17.3 −2.4 −1.8 0.2 1.5 Prob 0.017 0.011 0.0000.259 0.182 0.119 TILLER GDUSHD GDUSLK POLWT POLWT TASBLS PCT GDU GDUVALUE VALUE SCORE Stat ABS ABS ABS ABS % MN ABS Mean 1 3.6 147.0 146.7131.3 75.2 8.3 Mean 2 5.1 144.5 145.6 239.1 135.5 8.6 Locs 17 51 52 1313 7 Reps 17 51 52 13 13 7 Diff 1.5 2.4 1.1 −107.9 −60.2 −0.3 Prob 0.2080.000 0.042 0.000 0.000 0.356 TASSZ PLTHT EARHT STAGRN STKLDG BRTSTKSCORE CM CM SCORE % NOT % NOT Stat ABS ABS ABS ABS ABS ABS Mean 1 4.4227.7 91.4 3.1 100.0 98.6 Mean 2 6.9 223.0 81.3 2.6 98.6 91.7 Locs 35 363 17 4 3 Reps 35 36 3 17 4 3 Diff −2.5 4.7 10.2 0.5 1.4 6.9 Prob 0.0000.008 0.321 0.303 0.391 0.135 SCTGRN TEXEAR EARMLD BARPLT GLFSPT NLFBLTSCORE SCORE SCORE % NOT SCORE SCORE Stat ABS ABS ABS ABS ABS ABS Mean 17.6 5.0 6.1 95.8 5.9 5.1 Mean 2 7.4 6.5 6.3 97.9 4.4 5.4 Locs 14 4 10 2719 7 Reps 14 4 10 27 19 7 Diff 0.3 −1.5 −0.2 −2.0 1.5 −0.2 Prob 0.2630.014 0.619 0.120 0.000 0.356 SLFBLT STWWLT ANTROT MDMCPX FUSERS GIBERSSCORE SCORE SCORE SCORE SCORE SCORE Stat ABS ABS ABS ABS ABS ABS Mean 16.8 6.0 2.8 4.3 6.0 8.0 Mean 2 3.7 6.0 2.4 3.6 6.0 9.0 Locs 6 1 8 5 15 1Reps 6 1 8 5 15 1 Diff 3.1 0.0 0.4 0.7 0.0 −1.0 Prob 0.001 — 0.195 0.2450.935 — DIPERS COMRST ECB1LF ECB2SC CLDTST CLDTST SCORE SCORE SCORESCORE PCT PCT Stat ABS ABS ABS ABS ABS % MN Mean 1 4.9 5.3 8.5 3.5 84.998.4 Mean 2 5.0 5.4 7.0 2.5 78.7 91.0 Locs 5 8 1 1 10 10 Reps 5 8 1 1 1010 Diff −0.1 −0.2 1.5 1.0 6.2 7.3 Prob 0.815 0.623 — — 0.027 0.025KSZDCD HD SMT ERTLDG STLPCN ERTLPN LRTLPN PCT % NOT % NOT % NOT % NOT %NOT Stat ABS ABS ABS ABS ABS ABS Mean 1 4.8 98.7 25.0 30.0 100.0 20.0Mean 2 7.7 98.3 13.6 25.0 40.0 100.0 Locs 10 7 1 2 1 1 Reps 10 7 1 2 1 1Diff −2.9 0.4 11.4 5.0 60.0 −80.0 Prob 0.008 0.469 — 0.500 — —

TABLE 2B PAIRED INBRED COMPARISON REPORT Variety #1: PH4CV Variety #2:PHR03 YIELD YIELD MST TSTWT EGRWTH ESTCNT TILLER GDUSHD GDUSLK BU/A 56#BU/A 56# PCT LB/BU SCORE COUNT PCT GDU GDU Stat ABS % MN ABS ABS ABS ABSABS ABS ABS Mean 1 108.1 128.0 19.3 56.8 5.2 21.2 3.4 146.2 146.1 Mean 286.5 101.6 20.7 58.2 5.2 18.7 3.1 150.7 154.5 Locs 29 29 33 6 50 39 3290 91 Reps 29 29 33 6 50 39 32 90 91 Diff 21.7 26.5 1.4 −1.4 0.0 2.5 0.3−4.5 −8.4 Prob 0.000 0.000 0.003 0.277 0.892 0.000 0.821 0.000 0.000POLWT POLWT TASBLS TASSZ PLTHT EARHT STAGRN STKLDG BRTSTK VALUE VALUESCORE SCORE CM CM SCORE % NOT % NOT Stat ABS % MN ABS ABS ABS ABS ABSABS ABS Mean 1 120.9 77.7 8.3 4.4 224.3 92.2 3.4 100.0 99.0 Mean 2 180.1114.4 8.0 6.0 227.2 96.2 4.7 100.0 100.0 Locs 10 10 7 64 66 7 23 5 2Reps 10 10 7 64 66 7 23 5 2 Diff −59.2 −36.7 0.3 −1.6 −2.9 −4.0 −1.3 0.0−1.0 Prob 0.008 0.009 0.569 0.000 0.057 0.174 0.013 1.000 0.500 SCTGRNEARSZ TEXEAR EARMLD BARPLT DRPEAR GLFSPT NLFBLT SLFBLT SCORE SCORE SCORESCORE % NOT % NOT SCORE SCORE SCORE Stat ABS ABS ABS ABS ABS ABS ABS ABSABS Mean 1 7.5 5.0 5.4 5.9 96.7 100.0 6.1 5.2 6.8 Mean 2 7.1 6.0 7.1 7.397.5 100.0 5.3 5.5 4.9 Locs 20 2 8 17 47 1 18 5 4 Reps 20 2 8 17 47 1 185 4 Diff 0.5 −1.0 −1.8 −1.4 −0.7 0.0 0.8 0.3 0.0 Prob 0.035 1.000 0.0020.000 0.424 — 0.008 0.070 0.022 STWWLT ANTROT MDMCPX FUSERS GIBERSDIPERS COMRST ECB1LF ECB2SC SCORE SCORE SCORE SCORE SCORE SCORE SCORESCORE SCORE Stat ABS ABS ABS ABS ABS ABS ABS ABS ABS Mean 1 6.0 2.6 4.45.6 9.0 6.3 5.4 8.5 3.5 Mean 2 4.5 3.6 2.9 6.8 9.0 6.5 5.6 7.0 4.0 Locs1 7 4 18 1 5 10 1 1 Reps 1 7 4 18 1 5 10 1 1 Diff 1.5 −1.0 1.5 −1.2 0.0−0.2 −0.2 1.5 −0.5 Prob — 0.116 0.154 0.019 — 0.178 0.565 — — CLDTSTCLDTST KSZDCD HD SMT ERTLDG STLPCN ERTLPN LRTLPN PCT PCT PCT % NOT % NOT% NOT % NOT % NOT Stat ABS % MN ABS ABS ABS ABS ABS ABS Mean 1 86.9100.0 4.8 98.5 41.3 30.0 80.0 20.0 Mean 2 86.9 99.9 4.1 98.3 60.8 65.020.0 100.0 Locs 14 14 14 6 2 2 1 1 Reps 14 14 14 6 2 2 1 1 Diff 0.0 0.00.7 0.2 −19.4 −35.0 60.0 −80.0 Prob 1.000 0.994 0.345 0.877 0.411 0.500— —

TABLE 2C PAIRED INBRED COMPARISON REPORT Variety #1: PH4CV Variety #2:PH1BC YIELD YIELD MST EGRWTH ESTCNT TILLER BU/A 56# BU/A 56# PCT SCORECOUNT PCT Stat ABS % MN ABS ABS ABS ABS Mean 1 131.6 118.2 17.5 5.2 21.33.4 Mean 2 107.2 96.9 17.6 5.5 20.9 1.6 Locs 4 4 7 51 40 32 Reps 4 4 751 40 32 Diff 24.4 21.3 0.1 −0.3 0.3 −1.8 Prob 0.129 0.128 0.911 0.0410.463 0.206 GDUSHD GDUSLK POLWT POLWT TASBLS TASSZ GDU GDU VALUE VALUESCORE SCORE Stat ABS ABS ABS % MN ABS ABS Mean 1 146.7 146.6 120.9 77.78.3 4.4 Mean 2 150.0 149.5 156.6 102.8 8.1 5.0 Locs 101 103 10 10 7 72Reps 101 103 10 10 7 72 Diff −3.3 −2.9 −35.7 −25.2 0.1 −0.7 Prob 0.0000.000 0.031 0.060 0.356 0.000 PLTHT EARHT STAGRN STKLDG BRTSTK SCTGRN CMCM SCORE % NOT % NOT SCORE Stat ABS ABS ABS ABS ABS ABS Mean 1 224.891.8 3.3 100.0 97.8 7.5 Mean 2 243.5 94.3 4.3 98.6 91.3 7.4 Locs 76 8 314 1 23 Reps 76 8 31 4 1 23 Diff −18.7 −2.5 −1.0 1.4 6.5 0.0 Prob 0.0000.631 0.008 0.391 — 0.824 EARSZ TEXEAR EARMLD BARPLT DRPEAR GLFSPT SCORESCORE SCORE % NOT % NOT SCORE Stat ABS ABS ABS ABS ABS ABS Mean 1 5.05.3 5.8 97.8 100.0 6.0 Mean 2 4.0 7.1 5.7 96.6 100.0 5.8 Locs 2 10 17 461 22 Reps 2 10 17 46 1 22 Diff 1.0 −1.8 0.1 1.2 0.0 0.1 Prob 1.000 0.0000.808 0.192 — 0.576 NLFBLT SLFBLT STWWLT ANTROT MDMCPX FUSERS SCORESCORE SCORE SCORE SCORE SCORE Stat ABS ABS ABS ABS ABS ABS Mean 1 5.26.8 6.0 2.5 4.3 5.8 Mean 2 5.8 5.7 6.0 5.1 2.6 5.8 Locs 6 7 1 8 5 22Reps 6 7 1 8 5 22 Diff −0.7 1.1 0.0 −2.3 1.7 −0.0 Prob 0.318 0.006 —0.003 0.021 0.912 GIBERS DIPERS COMRST ECB1LF ECB2SC CLDTST SCORE SCORESCORE SCORE SCORE PCT Stat ABS ABS ABS ABS ABS ABS Mean 1 8.5 6.1 5.48.5 3.5 92.0 Mean 2 9.0 5.7 5.5 4.0 3.0 88.8 Locs 2 7 9 1 1 4 Reps 2 7 91 1 4 Diff −0.5 0.4 −0.1 4.5 0.5 3.3 Prob 0.500 0.253 0.892 — — 0.493CLDTST KSZDCD HD SMT ERTLDG ERTLPN PCT PCT % NOT % NOT % NOT Stat % MNABS ABS ABS ABS Mean 1 103.9 4.8 98.5 41.3 90.0 Mean 2 100.1 3.3 79.812.9 90.0 Locs 4 4 6 2 2 Reps 4 4 6 2 2 Diff 3.8 1.5 18.7 28.5 0.0 Prob0.475 0.476 0.171 0.250 1.000

TABLE 2D PAIRED INBRED COMPARISON REPORT Variety #1: PH4CV Variety #2:PH2EJ YIELD YIELD MST TSTWT EGRWTH ESTCNT TILLER GDUSHD GDUSLK BU/A 56#BU/A 56# PCT LB/BU SCORE COUNT PCT GDU GDU Stat ABS % MN ABS ABS ABS ABSABS ABS ABS Mean 1 104.5 129.9 19.6 56.6 5.3 21.2 3.6 146.9 147.1 Mean 279.5 98.3 16.7 57.1 4.9 20.4 4.2 152.9 154.4 Locs 25 25 26 7 51 39 29102 105 Reps 25 25 26 7 51 39 29 102 105 Diff 25.0 31 .7 −2.8 −0.5 0.40.8 0.5 −6.0 −7.2 Pro 0.000 0.000 0.000 0.580 0.004 0.139 0.552 0.0000.000 POLWT POLWT TASBLS TASSZ PLTHT EARHT STAGRN STKLDG BRTSTK VALUEVALUE SCORE SCORE CM CM SCORE % NOT % NOT Stat ABS % MN ABS ABS ABS ABSABS ABS ABS Mean 1 131.3 75.2 8.3 4.3 225.0 91.7 3.5 100.0 98.6 Mean 2131.0 75.6 8.1 2.8 234.4 105.6 2.3 100.0 95.8 Locs 13 13 7 73 77 9 32 53 Reps 13 13 7 73 77 9 32 5 3 Diff 0.3 −0.4 0.1 1.5 −9.4 −13.8 1.3 0.02.7 Prob 0.977 0.946 0.356 0.000 0.000 0.000 0.000 1.000 0.243 SCTGRNEARSZ TEXEAR EARMLD BARPLT DRPEAR GLFSPT NLFBLT SLFBLT SCORE SCORE SCORESCORE % NOT % NOT SCORE SCORE SCORE Stat ABS ABS ABS ABS ABS ABS ABS ABSABS Mean 1 7.5 5.0 5.5 6.1 97.2 100.0 6.0 5.2 6.8 Mean 2 7.2 5.0 5.9 6.797.3 100.0 3.9 5.0 4.3 Locs 25 2 10 17 51 1 24 6 6 Reps 25 2 10 17 51 124 6 6 Diff 0.3 0.0 −0.4 −0.6 −0.1 0.0 2.1 0.2 2.6 Prob 0.058 1.0000.309 0.116 0.898 — 0.000 0.695 0.001 STWWLT ANTROT MDMCPX FUSERS GIBERSDIPERS COMRST ECB1LF ECB2SC SCORE SCORE SCORE SCORE SCORE SCORE SCORESCORE SCORE Stat ABS ABS ABS ABS ABS ABS ABS ABS ABS Mean 1 6.0 2.8 4.36.0 8.7 6.1 5.7 8.5 3.5 Mean 2 7.0 3.3 3.2 6.9 9.0 6.5 6.9 7.0 2.0 Locs1 8 5 23 3 7 6 1 1 Reps 1 8 5 23 3 7 6 1 1 Diff −1.0 −0.5 1.1 −0.9 −0.3−0.4 −1.3 1.5 1.5 Prob — 0.425 0.130 0.007 0.423 0.078 0.081 — — CLDTSTCLDTST KSZDCD HD SMT ERTLDG STLPCN ERTLPN LRTLPN PCT PCT PCT % NOT % NOT% NOT % NOT % NOT Stat ABS % MN ABS ABS ABS ABS ABS ABS Mean 1 84.9 98.44.8 98.5 57.7 30.0 90.0 20.0 Mean 2 89.0 102.8 8.1 97.3 4.0 40.0 95.080.0 Locs 10 10 10 6 1 2 2 1 Reps 10 10 10 6 1 2 2 1 Diff −4.1 −4.4 −3.31.1 53.7 −10.0 −5.0 −60.0 Prob 0.267 0.333 0.008 0.588 — 0.500 0.500 —

TABLE 3A INBREDS IN HYBRID COMBINATION REPORT Variety #1: HYBRIDCONTAINING PH4CV Variety #2: 33J56 YIELD YIELD MST EGRWTH ESTCNT GDUSHDBU/A 56# BU/A 56# PCT SCORE COUNT GDU Stat ABS % MN % MN % MN % MN % MNMean 1 196.5 102.9 102.2 91.8 102.0 101.4 Mean 2 187.2 98.2 89.9 107.798.2 98.0 Locs 110 110 112 21 4 21 Reps 110 110 112 21 4 21 Diff 9.2 4.7−12.3 −15.9 3.8 3.3 Prob 0.000 0.000 0.000 0.005 0.160 0.000 GDUSLKSTKCNT PLTHT EARHT STAGRN ERTLSC GDU COUNT IN IN SCORE SCORE Stat % MN %MN % MN % MN % MN ABS Mean 1 102.0 100.0 95.8 90.1 117.4 6.0 Mean 2 98.1100.4 98.9 96.6 90.5 8.0 Locs 16 182 24 24 32 1 Reps 16 182 24 24 32 1Diff 3.9 −0.4 −3.1 −6.5 26.9 −2.0 Prob 0.003 0.423 0.000 0.011 0.000 —LRTLSC STKLDS STKLDG STKLDL EBTSTK ABTSTK SCORE SCORE % NOT % NOT % NOT% NOT Stat ABS ABS % MN % MN % MN % MN Mean 1 8.0 7.1 104.9 118.2 104.3129.1 Mean 2 3.0 5.1 98.3 86.5 101.8 107.0 Locs 1 14 8 14 2 8 Reps 1 148 14 2 8 Diff 5.0 2.0 6.6 31.7 2.4 22.0 Prob — 0.035 0.021 0.017 0.0830.047 TSTWT GLFSPT NLFBLT SLFBLT ANTROT CLN LB/BU SCORE SCORE SCORESCORE SCORE Stat ABS ABS ABS ABS ABS ABS Mean 1 56.9 5.6 4.8 6.7 3.9 4.0Mean 2 56.9 5.2 6.0 5.7 3.0 3.8 Locs 77 7 3 6 9 2 Reps 77 7 3 6 9 2 Diff0.1 0.4 −1.2 1.0 0.9 0.3 Prob 0.769 0.609 0.192 0.119 0.041 0.500 MDMCPXFUSERS DIPERS COMRST ECB1LF ECB2SC SCORE SCORE SCORE SCORE SCORE SCOREStat ABS ABS ABS ABS ABS ABS Mean 1 3.5 6.1 3.0 5.0 5.5 5.0 Mean 2 3.23.9 4.5 5.0 4.4 3.9 Locs 3 8 1 1 3 5 Reps 3 8 1 1 3 5 Diff 0.3 2.2 −1.50.0 1.1 1.0 Prob 0.423 0.001 — — 0.338 0.117 HSKCVR BRTSTK HD SMT ERTLPNLRTLPN STLPCN SCORE % NOT % NOT % NOT % NOT % NOT Stat ABS ABS ABS ABSABS ABS Mean 1 6.1 100.0 96.8 94.0 100.0 87.7 Mean 2 4.3 98.3 97.9 76.585.0 61.7 Locs 15 1 6 5 2 26 Reps 15 1 6 5 2 26 Diff 1.9 1.7 −1.1 17.515.0 26.0 Prob 0.000 — 0.359 0.245 0.500 0.000

TABLE 3B INBREDS IN HYBRID COMBINATION REPORT Variety #1: HYBRIDCONTAINING PH4CV Variety #2: 3335 YIELD YIELD MST EGRWTH STKCNT PLTHTBU/A 56# BU/A 56# PCT SCORE COUNT IN Stat ABS % MN % MN % MN % MN % MNMean 1 180.3 107.2 99.3 103.6 99.4 96.1 Mean 2 161.7 95.4 89.3 97.9102.0 95.5 Locs 21 21 21 3 23 3 Reps 21 21 21 3 23 3 Diff 18.7 11.7−10.0 5.7 −2.6 0.6 Prob 0.001 0.000 0.000 0.423 0.123 0.842 EARHT STAGRNSTKLDS STKLDG STKLDL DRPEAR IN SCORE SCORE % NOT % NOT % NOT Stat % MN %MN ABS % MN % MN % MN Mean 1 93.3 101.4 7.7 99.5 134.3 100.1 Mean 2 89.184.7 4.3 103.4 124.6 100.1 Locs 3 6 3 2 1 1 Reps 3 6 3 2 1 1 Diff 4.216.7 3.3 4.0 9.7 0.0 Prob 0.327 0.358 0.300 0.500 — — TSTWT GLFSPTSLFBLT ECB2SC HSKCVR LB/BU SCORE SCORE SCORE SCORE Stat ABS ABS ABS ABSABS Mean 1 57.3 5.0 7.0 5.0 3.5 Mean 2 57.4 5.0 5.0 5.0 3.5 Locs 14 1 11 2 Reps 14 1 1 1 2 Diff −0.1 0.0 2.0 0.0 0.0 Prob 0.736 — — — 1.000

TABLE 3C INBREDS IN HYBRID COMBINATION REPORT Variety #1: HYBRIDCONTAINING PH4CV Variety #2: 3245 YIELD YIELD MST EGRWTH ESTCNT GDUSHDGDUSLK STKCNT BU/A 56# BU/A 56# PCT SCORE COUNT GDU GDU COUNT Stat ABS %MN % MN % MN % MN % MN % MN % MN Mean 1 188.2 104.9 102.3 97.2 102.0100.5 100.3 99.4 Mean 2 174.3 96.9 94.3 108.1 101.9 99.5 101.6 100.7Locs 77 77 79 13 4 8 7 103 Reps 77 77 79 13 4 8 7 103 Diff 13.9 8.0 −8.0−10.8 0.1 1.0 −1.3 −1.3 Prob 0.000 0.000 0.000 0.038 0.962 0.058 0.0680.085 PLTHT EARHT STAGRN ERTLSC LRTLSC STKLDS STKLDG STKLDL IN IN SCORESCORE SCORE SCORE % NOT % NOT Stat % MN % MN % MN ABS ABS ABS % MN % MNMean 1 95.9 91.2 112.9 6.0 8.0 7.1 103.9 105.4 Mean 2 98.0 94.5 78.2 9.07.0 6.6 101.5 103.8 Locs 13 13 22 1 1 18 8 8 Reps 13 13 22 1 1 18 8 8Diff −2.1 −3.3 34.8 −3.0 1.0 0.5 2.4 1.6 Prob 0.023 0.135 0.000 — —0.509 0.306 0.825 EBTSTK ABTSTK TSTWT GLFSPT NLFBLT SLFBLT ANTROT CLN %NOT % NOT LB/BU SCORE SCORE SCORE SCORE SCORE Stat % MN % MN ABS ABS ABSABS ABS ABS Mean 1 104.3 108.6 57.2 5.0 5.0 7.0 3.6 4.0 Mean 2 102.289.2 58.4 4.1 4.0 4.3 3.2 3.5 Locs 2 3 56 4 2 2 6 1 Reps 2 3 56 4 2 2 61 Diff 2.1 19.4 −1.1 0.9 1.0 2.8 0.4 0.5 Prob 0.500 0.227 0.000 0.1330.295 0.170 0.392 MDMCPX FUSERS DIPERS COMRST ECB1LF ECB2SC HSKCVR HDSMT SCORE SCORE SCORE SCORE SCORE SCORE SCORE % NOT Stat ABS ABS ABS ABSABS ABS ABS ABS Mean 1 3.0 5.5 3.0 5.0 6.3 4.5 4.6 96.1 Mean 2 2.0 4.34.0 3.0 5.3 4.3 5.4 94.0 Locs 1 3 1 1 2 3 8 3 Reps 1 3 1 1 2 3 8 3 Diff1.0 1.2 −1.0 2.0 1.0 0.2 −0.8 2.1 Prob — 0.250 — — 0.295 0.808 0.0800.614

TABLE 3D INBREDS IN HYBRID COMBINATION REPORT Variety #1: HYBRIDCONTAINING PH4CV Variety #2: 32H58 YIELD YIELD MST EGRWTH GDUSHD GDUSLKBU/A 56# BU/A 56# PCT SCORE GDU GDU Stat ABS % MN % MN % MN % MN % MNMean 1 201.6 101.8 100.8 88.7 101.9 103.3 Mean 2 192.4 97.4 95.1 106.498.2 101.8 Locs 53 53 53 11 13 9 Reps 53 53 53 11 13 9 Diff 9.3 4.4 −5.7−17.7 3.7 1.5 Prob 0.007 0.018 0.000 0.002 0.181 0.003 STKCNT PLTHTEARHT STAGRN STKLDG STKLDL COUNT IN IN SCORE % NOT % NOT Stat % MN % MN% MN % MN % MN % MN Mean 1 100.5 95.6 89.6 119.5 109.4 135.0 Mean 2 99.395.7 96.9 95.3 64.3 93.6 Locs 95 13 13 15 1 7 Reps 95 13 13 15 1 7 Diff1.2 −0.1 −7.3 24.2 45.0 41.4 Prob 0.023 0.977 0.002 0.017 — 0.052 ABTSTKTSTWT GLFSPT NLFBLT SLFBLT ANTROT % NOT LB/BU SCORE SCORE SCORE SCOREStat % MN ABS ABS ABS ABS ABS Mean 1 141.4 56.7 6.0 4.5 6.6 4.5 Mean 2108.4 57.3 5.9 4.0 4.5 4.0 Locs 5 35 4 1 5 3 Reps 5 35 4 1 5 3 Diff 33.0−0.7 0.1 0.5 2.1 0.5 Prob 0.075 0.038 0.824 — 0.006 1.000 CLN MDMCPXFUSERS ECB1LF ECB2SC HSKCVR SCORE SCORE SCORE SCORE SCORE SCORE Stat ABSABS ABS ABS ABS ABS Mean 1 4.0 3.8 6.4 4.0 5.4 6.9 Mean 2 3.0 3.0 4.53.3 4.9 5.6 Locs 1 2 5 1 3 9 Reps 1 2 5 1 3 9 Diff 1.0 0.8 1.9 0.7 0.61.3 Prob — 0.205 0.095 — 0.525 0.016 BRTSTK HD SMT ERTLPN LRTLPN STLPCN% NOT % NOT % NOT % NOT % NOT Stat ABS ABS ABS ABS ABS Mean 1 100.0 97.594.0 100.0 87.7 Mean 2 100.0 97.1 72.0 100.0 76.0 Locs 1 3 5 2 26 Reps 15 5 2 26 Diff 0.0 0.4 22.0 0.0 11.7 Prob — 0.877 0.051 1.000 0.020

TABLE 3E INBREDS IN HYBRID COMBINATION REPORT Variety #1: HYBRIDCONTAINING PH4CV Variety #2: 31G98 YIELD YIELD MST EGRWTH ESTCNT GDUSHDBU/A 56# BU/A 56# PCT SCORE COUNT GDU Stat ABS % MN % MN % MN % MN % MNMean 1 196.7 103.2 102.2 91.4 102.0 101.0 Mean 2 195.2 103.1 100.1 94.196.8 102.9 Locs 117 117 119 22 4 26 Reps 117 117 119 22 4 26 Diff 1.50.1 −2.2 −2.7 5.2 −1.8 Prob 0.507 0.914 0.004 0.430 0.090 0.000 GDUSLKSTKCNT PLTHT EARHT STAGRN ERTLSC GDU COUNT IN IN SCORE SCORE Stat % MN %MN % MN % MN % MN ABS Mean 1 101.5 100.1 95.8 90.4 116.6 6.0 Mean 2103.0 99.6 101.5 105.2 108.8 9.0 Locs 20 194 28 28 35 1 Reps 20 194 2828 35 1 Diff −1.5 0.6 −5.6 −14.8 7.8 −3.0 Prob 0.006 0.230 0.000 0.0000.175 — LRTLSC STKLDS STKLDG STKLDL EBTSTK ABTSTK SCORE SCORE % NOT %NOT % NOT % NOT Stat ABS ABS % MN % MN % MN % MN Mean 1 8.0 7.1 104.9115.9 104.3 129.1 Mean 2 4.0 6.6 96.2 106.5 102.0 54.9 Locs 1 14 8 18 28 Reps 1 14 8 18 2 8 Diff 4.0 0.5 8.7 9.4 2.3 74.2 Prob — 0.561 0.0480.324 0.602 0.000 TSTWT GLFSPT NLFBLT SLFBLT ANTROT CLN LB/BU SCORESCORE SCORE SCORE SCORE Stat ABS ABS ABS ABS ABS ABS Mean 1 56.9 5.6 4.86.7 3.9 4.0 Mean 2 56.1 5.1 5.7 4.2 5.1 4.3 Locs 84 7 3 7 10 2 Reps 84 73 7 10 2 Diff 0.8 0.4 −0.8 2.5 −1.2 −0.3 Prob 0.000 0.289 0.300 0.0000.055 0.500 MDMCPX FUSERS DIPERS COMRST ECB1LF ECB2SC SCORE SCORE SCORESCORE SCORE SCORE Stat ABS ABS ABS ABS ABS ABS Mean 1 3.5 6.1 3.0 5.05.5 5.0 Mean 2 3.2 4.9 5.0 5.0 5.3 4.1 Locs 3 8 1 1 3 5 Reps 3 8 1 1 3 5Diff 0.3 1.2 −2.0 0.0 0.2 0.9 Prob 0.529 0.029 — — 0.423 0.276 HSKCVRBRTSTK HD SMT ERTLPN LRTLPN STLPCN SCORE % NOT % NOT % NOT % NOT % NOTStat ABS ABS ABS ABS ABS ABS Mean 1 6.0 100.0 96.8 84.6 100.0 88.9 Mean2 5.3 98.2 96.8 83.5 100.0 80.7 Locs 20 1 6 8 2 30 Reps 20 1 6 8 2 30Diff 0.7 1.8 −0.0 1.0 0.0 8.2 Prob 0.010 — 0.980 0.934 1.000 0.002Genetic Marker Profile through SSR

The present invention comprises an inbred corn plant which ischaracterized by the molecular and physiological data presented hereinand in the representative sample of said line deposited with the ATCC.Further provided by the invention is a hybrid corn plant formed by thecombination of the disclosed inbred corn plant or plant cell withanother corn plant or cell and characterized by being heterozygous forthe molecular data of the inbred.

In addition to phenotypic observations, a plant can also be identifiedby its genotype. The genotype of a plant can be characterized through agenetic marker profile which can identify plants of the same variety ora related variety or be used to determine or validate a pedigree.Genetic marker profiles can be obtained by techniques such asRestriction Fragment Length Polymorphisms (RFLPs), Randomly AmplifiedPolymorphic DNAs (RAPDs), Arbitrarily Primed Polymerase Chain Reaction(AP-PCR), DNA Amplification Fingerprinting (DAF), Sequence CharacterizedAmplified Regions (SCARs), Amplified Fragment Length Polymorphisms(AFLPs), Simple Sequence Repeats (SSRs) which are also referred to asMicrosatellites, and Single Nucleotide Polymorphisms (SNPs). Forexample, see Berry, Don, et al., “Assessing Probability of AncestryUsing Simple Sequence Repeat Profiles: Applications to Maize Hybrids andInbreds”, Genetics, 2002, 161:813-824, which is incorporated byreference herein in its entirety.

Particular markers used for these purposes are not limited to the set ofmarkers disclosed herein, but are envisioned to include any type ofmarker and marker profile which provides a means of distinguishingvarieties. In addition to being used for identification of Inbred LinePH4CV, a hybrid produced through the use of PH4CV, and theidentification or verification of pedigree for progeny plants producedthrough the use of PH4CV, the genetic marker profile is also useful inbreeding and developing single gene conversions.

Means of performing genetic marker profiles using SSR polymorphisms arewell known in the art. SSRs are genetic markers based on polymorphismsin repeated nucleotide sequences, such as microsatellites. A markersystem based on SSRs can be highly informative in linkage analysisrelative to other marker systems in that multiple alleles may bepresent. Another advantage of this type of marker is that, through useof flanking primers, detection of SSRs can be achieved, for example, bythe polymerase chain reaction (PCR), thereby eliminating the need forlabor-intensive Southern hybridization. The PCR™ detection is done byuse of two oligonucleotide primers flanking the polymorphic segment ofrepetitive DNA. Repeated cycles of heat denaturation of the DNA followedby annealing of the primers to their complementary sequences at lowtemperatures, and extension of the annealed primers with DNA polymerase,comprise the major part of the methodology.

Following amplification, markers can be scored by gel electrophoresis ofthe amplification products. Scoring of marker genotype is based on thesize of the amplified fragment as measured by molecular weight (MW)rounded to the nearest integer. While variation in the primer used or inlaboratory procedures can affect the reported molecular weight, relativevalues should remain constant regardless of the specific primer orlaboratory used. When comparing lines it is preferable if all SSRprofiles are performed in the same lab. The SSR analyses reported hereinwere conducted in-house at Pioneer Hi-Bred. An SSR service is availableto the public on a contractual basis by Paragen (formerly Celera AgGen)in Research Triangle Park, N.C.

Primers used for the SSRs reported herein are publicly available and maybe found at the World Wide Web at agron.missouri.edu/maps.html(sponsored by the University of Missouri), in Sharopova et al. (PlantMol. Biol. 48(5-6):463-481), Lee et al (Plant Mol. Biol. 48(5-6);453-461), or reported herein. Some marker information may be availablefrom Paragen.

Map information is provided in centimorgans (cM) and based on acomposite map developed by Pioneer Hi-Bred. This composite map wascreated by identifying common markers between various maps and usinglinear regression to place the intermediate markers. The reference mapused was UMC98. Map positions for the SSR markers reported herein willvary depending on the mapping population used. Any chromosome numbersreported in parenthesis represent other chromosome locations for suchmarker that have been reported in the literature or on the Maize DB. Mappositions are available on the Maize DB for a variety of differentmapping populations.

TABLE 4 SSR Profile Locus Chrom # Position PH4CV mwt PHI056 1 4.25 252PHI427913 1 26.71 131 BNLG1627 1 34.03 201 BNLG1127 1 38.07 100 BNLG19531 38.34 203 BNLG1484 1 53.32 148 PHI339017 1 70.05 148 BNLG1886 1 91.64144 BNLG1057 1 142.56 253 PHI002 1 168.21 72 PHI335539 1 178.84 91BNLG1331 1 193.48 124 BNLG1597 1 199.42 191 PHI308707 1 211.59 134PHI265454 1 221.46 221 PHI064 1 243.08 88 PHI109275 1 unknown 128BNLG1832 1 unknown 218 BNLG1083 1 unknown 223 BNLG1016 1 unknown 259PHI96100 2 6.99 284 BNLG1064 2 64.21 191 PHI109642 2 69.93 134 BNLG10182 77.92 141 BNLG1138 2 121.18 227 PHI328189 2 145.57 124 PHI251315 2149.77 127 PHI101049 2 207.93 230 PHI402893 2 unknown 221 PHI090 2unknown 141 PHI083 2 unknown 132 BNLG1831 2 unknown 193 PHI453121 3 0.2216 PHI104127 3 5.68 165 BNLG1144 3 23.52 118 BNLG1647 3 33.98 154PHI374118 3 53.66 227 BNLG1452 3 58.58 128 BNLG1113 3 58.65 137 PHI053 367.9 169 PHI102228 3 104.98 131 BNLG1951 3 108.98 130 BNLG1160 3 110.2225 BNLG2241 3 unknown 119 BNLG1035 3 unknown 104 PHI213984 4 25.02 287PHI096 4 61.84 238 PHI079 4 65.46 188 BNLG1937 4 65.49 241 BNLG1265 467.31 232 BNLG1189 4 108.12 133 BNLG2244 4 122.51 202 PHI093 4 126.18284 PHI314704 4 159.67 139 PHI438301 4 819.88 210 PHI072 4 unknown 143BNLG1006 5 15.9 219 PHI109188 5 77.97 170 BNLG653 5 88.43 154 BNLG1208 594.95 121 PHI386223 5 95.46 141 PHI330507 5 102.23 135 PHI333597 5103.48 219 PHI196387 5 133.31 231 PHI085 5 136.05 252 BNLG1118 5 149.5384 BNLG1711 5 178.37 183 PHI423796 6 31.29 131 PHI389203 6 83.56 307BNLG1041 6 98.1 197 BNLG1174 6 99.41 222 PHI445613 6 106.74 100PHI364545 6 126.24 138 PHI070 6 129.9 83 BNLG1759 6 129.9 151 PHI034 754.75 141 BNLG2271 7 95.38 237 PHI260485 7 134.62 288 PHI069 7 137.5 203PHI116 7 149.22 172 PHI420701 8 24.32 294 BNLG1194 8 27.24 195 BNLG20828 55.3 174 PHI115 8 64.03 303 BNLG2046 8 75.05 327 BNLG1176 8 79.72 221BNLG1152 8 111.61 161 BNLG1065 8 127.89 219 BNLG1056 8 193.84 111 PHI0158 210.92 85 BNLG2122 9 32.11 222 PHI448880 9 126.07 188 PHI236654 9157.45 120 PHI108411 9 169.83 121 PHI033 9 unknown 252 PHI041 10 9.6 205PHI059 10 40.55 156 PHI96342 10 52.8 249 BNLG1079 10 62.12 172 PHI050 1063 86 PHI062 10 69.31 164 BNLG1074 10 85.74 174 PHI301654 10 91.26 132PHI323152 10 117.1 137 BNLG1450 10 126.36 243 BNLG1185 10 142.38 154

The SSR profile of Inbred PH4CV can be used to identify hybridscomprising PH4CV as a parent, since such hybrids will comprise the samealleles as PH4CV. Because an inbred is essentially homozygous at allrelevant loci, an inbred should, in almost all cases, have only oneallele at each locus. In contrast, a genetic marker profile of a hybridshould be the sum of those parents, e.g., if one inbred parent had theallele 168 (base pairs) at a particular locus, and the other inbredparent had 172 the hybrid is 168.172 (heterozygous) by inference.Subsequent generations of progeny produced by selection and breeding areexpected to be of genotype 168 (homozygous), 172 (homozygous), or168.172 for that locus position. When the F1 plant is used to produce aninbred, the locus should be either 168 or 172 for that position.

In addition, plants and plant parts substantially benefiting from theuse of PH4CV in their development such as PH4CV comprising a single geneconversion, transgene, or genetic sterility factor, may be identified byhaving a molecular marker profile with a high percent identity to PH4CV.Such a percent identity might be 98%, 99%, 99.5% or 99.9% identical toPH4CV.

The SSR profile of PH4CV also can be used to identify essentiallyderived varieties and other progeny lines developed from the use ofPH4CV, as well as cells and other plant parts thereof. Progeny plantsand plant parts produced using PH4CV may be identified by having amolecular marker profile of at least 25%, 30%, 35%, 40%, 45%, 50%, 55%,60%, 65%, 70%, 75%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 99.5% geneticcontribution from inbred line PH4CV.

Deposits

Applicant has made a deposit of at least 2500 seeds of Inbred Maize LinePH4CV with the American Type Culture Collection (ATCC), Manassas, Va.20110 USA, ATCC Deposit No. PTA-4673. The seeds deposited with the ATCCon Sep. 18, 2002 were taken from the deposit maintained by PioneerHi-Bred International, Inc., 800 Capital Square, 400 Locust Street, DesMoines, Iowa 50309-2340 since prior to the filing date of thisapplication. Access to this deposit will be available during thependency of the application to the Commissioner of Patents andTrademarks and persons determined by the Commissioner to be entitledthereto upon request. Upon allowance of any claims in the application,the Applicant will make the deposit available to the public pursuant to37 C.F.R. § 1.808. This deposit of the Inbred Maize Line PH4CV will bemaintained in the ATCC depository, which is a public depository, for aperiod of 30 years, or 5 years after the most recent request, or for theenforceable life of the patent, whichever is longer, and will bereplaced if it becomes nonviable during that period. Additionally,Applicant has satisfied all the requirements of 37 C.F.R. §§1.801-1.809,including providing an indication of the viability of the sample upondeposit. Applicant has no authority to waive any restrictions imposed bylaw on the transfer of biological material or its transportation incommerce. Applicant does not waive any infringement of his rightsgranted under this patent or under the Plant Variety Protection Act (7USC 2321 et seq.). U.S. Plant Variety Protection of Inbred Maize LinePH4CV has been applied for under Application No. 200200176.

All publications, patents and patent applications mentioned in thespecification are indicative of the level of those skilled in the art towhich this invention pertains. All such publications, patents and patentapplications are incorporated by reference herein to the same extent asif each was specifically and individually indicated to be incorporatedby reference herein.

The foregoing invention has been described in detail by way ofillustration and example for purposes of clarity and understanding.However, it will be obvious that certain changes and modifications suchas single gene conversions and mutations, somocional variants, variantindividuals selected from large populations of the plants of the instantinbred and the like may be practiced within the scope of the Invention,as limited only by the scope of the appended claims.

1. A seed of maize inbred line designated PH4CV, representative seed ofsaid line having been deposited under ATCC Accession No. PTA-4673.
 2. Amaize plant, or a part thereof, produced by growing the seed of claim 1.3. The maize plant of claim 2, wherein said plant has been detasseled.4. A tissue culture of regenerable cells produced from the plant ofclaim
 2. 5. A protoplast produced from the tissue culture of claim
 4. 6.The tissue culture of claim 4, wherein cells of the tissue culture areproduced from a tissue selected from the group consisting of leaf,pollen, embryo, root, root tip, anther, silk, flower, kernel, ear, cob,husk and stalk.
 7. A maize plant regenerated from the tissue culture ofclaim 4, said plant having all the morphological and physiologicalcharacteristics of inbred line PH4CV, representative seed of said inbredline having been deposited under ATCC Accession No. PTA-4673.
 8. Amethod for producing an F1 hybrid maize seed, comprising crossing theplant of claim 2 with a different maize plant and harvesting theresultant F1 hybrid maze seed.
 9. A method of producing a male sterilemaize plant comprising transforming the maize plant of claim 2 with anucleic acid molecule that confers male sterility.
 10. A male sterilemaize plant produced by the method of claim
 9. 11. A method of producingan herbicide resistant maize plant comprising transforming the maizeplant of claim 2 with a transgene that confers herbicide resistance. 12.An herbicide resistant maize plant produced by the method of claim 11.13. The maize plant of claim 12, wherein the transgene confersresistance to an herbicide selected from the group consisting ofimidazolinone, sulfonylurea, glyphosate, glufosinate,L-phosphinothricin, triazine and benzonitrile.
 14. A method of producingan insect resistant maize plant comprising transforming the maize plantof claim 2 with a transgene that confers insect resistance.
 15. Aninsect resistant maize plant produced by the method of claim
 14. 16. Themaize plant of claim 15, wherein the transgene encodes a Bacillusthuringiensis endotoxin.
 17. A method of producing a disease resistantmaize plant comprising transforming the maize plant of claim 2 with atransgene that confers disease resistance.
 18. A disease resistant maizeplant produced by the method of claim
 17. 19. A method of producing amaize plant with decreased phytate content comprising transforming themaize plant of claim 2 with a transgene encoding phytase.
 20. A maizeplant with decreased phytate content produced by the method of claim 19.21. A method of producing a maize plant with modified fatty acidmetabolism or modified carbohydrate metabolism comprising transformingthe maize plant of claim 2 with a transgene encoding a protein selectedfrom the group consisting of fructosyltransferase, levansucrase,alpha-amylase, invertase and starch branching enzyme or encoding anantisense of stearyl-ACP desaturase.
 22. A maize plant, with modifiedfatty acid metabolism or modified carbohydrate metabolism, produced bythe method of claim
 21. 23. The maize plant of 22 wherein the transgeneconfers a trait selected from the group consisting of waxy starch andincreased amylose starch.
 24. A maize plant, or part thereof, having allthe physiological and morphological characteristics of the inbred linePH4CV, representative seed of said inbred line having been depositedunder ATCC Accession No. PTA-4673.
 25. A method of introducing a desiredtrait into maize inbred line PH4CV comprising: (a) crossing PH4CV plantsgrown from PH4CV seed, representative seed of which has been depositedunder ATCC Accession No. PTA-4673, with plants of another maize linethat comprise a desired trait to produce F1 progeny plants, wherein thedesired trait is selected from the group consisting of male sterility,herbicide resistance, insect resistance, disease resistance and waxystarch; (b) selecting F1 progeny plants that have the desired trait toproduce selected F1 progeny plants; (c) crossing the selected progenyplants with the PH4CV plants to produce backcross progeny plants; (d)selecting for backcross progeny plants that have the desired trait andphysiological and morphological characteristics of maize inbred linePH4CV listed in Table 1 to produce selected backcross progeny plants;and (e) repeating steps (c) and (d) three or more times in succession toproduce selected fourth or higher backcross progeny plants that comprisethe desired trait and all of the physiological and morphologicalcharacteristics of maize inbred line PH4CV listed in Table 1 asdetermined at the 5% significance level when grown in the sameenvironmental conditions.
 26. A plant produced by the method of claim25, wherein the plant has the desired trait and all of the physiologicaland morphological characteristics of maize inbred line PH4CV listed inTable 1 as determined at the 5% significance level when grown in thesame environmental conditions.
 27. The plant of claim 26 wherein thedesired trait is herbicide resistance and the resistance is conferred toan herbicide selected from the group consisting of imidazolinone,sulfonylurea, glyphosate, glufosinate, L-phosphinothricin, triazine andbenzonitrile.
 28. The plant of claim 26 wherein the desired trait isinsect resistance and the insect resistance is conferred by a transgeneencoding a Bacillus thuringiensis endotoxin.
 29. The plant of claim 26wherein the desired trait is male sterility and the trait is conferredby a cytoplasmic nucleic acid molecule that confers male sterility. 30.A method of modifying fatty acid metabolism, phytic acid metabolism orcarbohydrate in maize inbred line PH4CV comprising: (a) crossing PH4CVplants grown from PH4CV seed, representative seed of which has beendeposited under ATCC Accession No. PTA-4673, with plants of anothermaize line that comprise a nucleic acid molecule encoding an enzymeselected from the group consisting of phytase, fructosyltransferase,levansucrase, alpha-amylase, invertase and starch branching enzyme orencoding an antisense of stearyl-ACP desaturase; (b) selecting F1progeny plants that have said nucleic acid molecule to produce selectedF1 progeny plants; (c) crossing the selected progeny plants with thePH4CV plants to produce backcross progeny plants; (d) selecting forbackcross progeny plants that have said nucleic acid molecule andphysiological and morphological characteristics of maize inbred linePH4CV listed in Table 1 to produce selected backcross progeny plants;and (e) repeating steps (c) and (d) three or more times in succession toproduce selected fourth or higher backcross progeny plants that comprisesaid nucleic acid molecule and have all of the physiological andmorphological characteristics of maize inbred line PH4CV listed in Table1 as determined at the 5% significance level when grown in the sameenvironmental conditions.
 31. A plant, with modified fatty acidmetabolism, modified phytic acid metabolism, or modified carbohydratemetabolism, produced by the method of claim 30, wherein the plantcomprises the nucleic acid molecule and has all of the physiological andmorphological characteristics of maize inbred line PH4CV listed in Table1 as determined at the 5% significance level when grown in the sameenvironmental conditions.